Team:UT Dallas/notebook data

From 2012.igem.org

(Difference between revisions)
Line 2: Line 2:
<div id="page1" class='notebook_page'>Cover page</div>
<div id="page1" class='notebook_page'>Cover page</div>
<div id="page2" class='notebook_page'><p>
<div id="page2" class='notebook_page'><p>
-
<strong>July 19</strong>
+
    June 6
</p>
</p>
 +
<br/>
<p>
<p>
-
Transformations
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; BBa_I721001: lead promoter
+
    Transformation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        BBa_J61002: “RFP” promoter
 +
    </li>
 +
    <li>
 +
        BBa_K081012: GFP protein generator (RBS-GFP)
 +
    </li>
 +
    <li>
 +
        BBa_J23103: constitutive family member
 +
    </li>
 +
    <li>
 +
        Bba_J04450: RFP Coding Device (pLas-RBS-RFP-Term)
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; BBa_K091109: LuxS
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; BBa_C0078: LasI
+
    June 7
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; BBa_C0062: LuxR
+
    Colonies picked and glycerol stocked made
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        RFP promoter: 5 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        RBS-GFP: 2 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        constitutive family promoter: 2 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        pLas-RBS-RFP-Term: 6 colonies seen: 1 colony picked
 +
    </li>
 +
</ul>
 +
<br/>
 +
</div>
 +
<div id="page3" class='notebook_page'>
<p>
<p>
-
&#9679; BBa_C0061: LuxI
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; BBa_E0432: RBS-YFP
+
    June 8
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>July 20</strong>
+
    Plasmid isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        RBS-GFP
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            Concentration:111.2 ng/ul
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        constututive family member
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            Concentration: 109.8 ng/ul
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        pLasI-RBS-RFP-Term
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            Concentration: 113.9 ng/ul
 +
        </li>
 +
    </ul>
 +
</ul>
 +
<br/>
 +
</div>
 +
<div id="page4" class='notebook_page'>
 +
<p>
<p>
-
Colonies picked
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: 20 colonies seen: 1 colony picked
+
    June 11
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxR: 5 colonies seen: 1 colony picked
+
    Digestion
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        RBS-GFP (E/S)---(wrong enzymes used)
 +
    </li>
 +
    <li>
 +
        constitutive promoter (E/X)---(wrong enzymes used)
 +
    </li>
 +
    <li>
 +
        constitutive promoter (S/P)
 +
    </li>
 +
    <li>
 +
        pLasI-RBS-RFP-Term (X/P)
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; LuxI: 5 colonies seen: 1 colony picked
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; RBS-YFP: no colonies seen
+
    June 12
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LasI: no colonies seen
+
    Digestion
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        RBS-GFP (X/P)
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; lead promoter: no colonies seen
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector: constitutive promoter
 +
    </li>
 +
    <li>
 +
        Insert: RBS-GFP
 +
    </li>
 +
    <li>
 +
        0:1--- 0 colonies counted
 +
    </li>
 +
    <li>
 +
        2:1--- 7 colonies counted
 +
    </li>
 +
</ul>
 +
<br/>
</div>
</div>
-
<div id="page3" class='notebook_page'><p>
+
<div id="page5" class='notebook_page'>
-
<strong>July 20</strong>
+
<p>
 +
    June 13
</p>
</p>
 +
<br/>
<p>
<p>
-
Repeat Transformation
+
    Incubate constitutive promoter-RBS-GFP in LB carb broth overnight @ 37*C and 220 rpm
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        2:1--- 2 colonies picked
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; lead promoter
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LasI
+
    June 14
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; RBS-YFP
+
    Glycerol stock and miniprep
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        constitutive promoter-RBS-GFP
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            colony 1 picked: 91 ng/ul
 +
        </li>
 +
        <li>
 +
            colony 2 picked: 97 ng/ul
 +
        </li>
 +
    </ul>
 +
</ul>
<p>
<p>
-
<strong>July 21</strong>
+
    Test Digestion for each plasmid: correct results received
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
Glycerol Stock
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS
+
    June 15
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxR
+
    Plasmid isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        prom-RBS-GFP: Concentration: 127 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
 +
</div>
 +
<div id="page6" class='notebook_page'>
 +
<p>
<p>
-
&#9679; LuxI
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Colonies picked
+
    July 16
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; RBS-YFP: 1 colony picked
+
    Transformations
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Bba_I763035 pTetR-RBS-LacI-RBS-GFP-Term-Term
 +
    </li>
 +
    <li>
 +
        Bba_K093005 RBS-RFP
 +
    </li>
 +
    <li>
 +
        Bba_R0010 pLacI
 +
    </li>
 +
    <li>
 +
        Bba_C0040 TetR
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; LasI: 1 colony picked
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; lead promoter: no colones seen
+
    July 17
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Bba_I763035 pTetR-RBS-LacI-RBS-GFP-Term-Term
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            2 colonies picked
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        Bba_K093005 RBS-RFP
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            2 colonies picked
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        Bba_R0010 pLacI
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            2 colonies picked
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        Bba_C0040 TetR
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            2 colonies picked
 +
        </li>
 +
    </ul>
 +
</ul>
 +
</div>
 +
<div id="page7" class='notebook_page'>
 +
 +
<br/>
<p>
<p>
-
Digestion &amp; Gel Extraction
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS (wrong enzymes used)
+
    July 18
</p>
</p>
-
</div>
+
<br/>
-
<div id="page4" class='notebook_page'><p>
+
<p>
-
&#9679; LuxR: Concentration: 11.1 ng/&#956;l
+
    Glycerol stocks made
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-RBS-LacI-RBS-GFP-Term-Term
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            Concentration: 236.2 ng/μl
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        RBS-RFP
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            Concentration: 92.9 ng/μl
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        pLacI
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            Concentration: 60.6 ng/μl
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        TetR
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            Concentration: 110.9 ng/μl
 +
        </li>
 +
    </ul>
 +
</ul>
<p>
<p>
-
&#9679; LuxI: Concentration: 7.1 ng/&#956;l
+
    Digestion
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-RBS-LacI-RBS-GFP-Term-Term (E/P)
 +
    </li>
 +
    <li>
 +
        RBS-RFP (X/P)
 +
    </li>
 +
    <li>
 +
        pLacI (S/P)
 +
    </li>
 +
    <li>
 +
        TetR (X/P)
 +
    </li>
 +
</ul>
 +
<br/>
</div>
</div>
-
<div id="page5" class='notebook_page'><p>
+
<div id="page8" class='notebook_page'>
-
<strong>July 22</strong>
+
 
 +
<p>
 +
    Gel Extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        RBS-RFP
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            Concentration: 10.8 ng/μl
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        pLac
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            Concentration: 19.6 ng/μl
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        TetR
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            Concentration: 6.2 ng/μl
 +
        </li>
 +
    </ul>
 +
</ul>
 +
<br/>
<p>
<p>
-
Plasmid Isolation
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector: pLac
 +
    </li>
 +
    <li>
 +
        Insert: TetR
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: Concentration: 75.2 ng/&#956;l
+
    Transformation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-TetR
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            0:1, 0 colonies seen
 +
        </li>
 +
        <li>
 +
            2:1, 20 colonies seen, 1 picked
 +
        </li>
 +
        <li>
 +
            4:1, 20 colonies seen, 1 picked
 +
        </li>
 +
    </ul>
 +
</ul>
 +
<br/>
 +
</div>
 +
<div id="page9" class='notebook_page'>
 +
<p>
<p>
-
&#9679; LasI: Concentration: 90.1 ng/&#956;l
+
    July 19
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; RBS-YFP: Concentration: 76.4 ng/&#956;l
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>July 23</strong>
+
    Transformations
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        BBa_I721001: lead promoter
 +
    </li>
 +
    <li>
 +
        BBa_K091109: LuxS
 +
    </li>
 +
    <li>
 +
        BBa_C0078: LasI
 +
    </li>
 +
    <li>
 +
        BBa_C0062: LuxR
 +
    </li>
 +
    <li>
 +
        BBa_C0061: LuxI
 +
    </li>
 +
    <li>
 +
        BBa_E0432: RBS-YFP
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
DIgestion and gel extraction
+
    July 20
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS (X/P) Concentration: not right size to be repeated
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LasI (E/S) Concentration: 20.3 ng/&#956;l
+
    Colonies picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS: 20 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        LuxR: 5 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        LuxI: 5 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        RBS-YFP: no colonies seen
 +
    </li>
 +
</div>
 +
<div id="page10" class='notebook_page'>
 +
 +
    <li>
 +
        LasI: no colonies seen
 +
    </li>
 +
    <li>
 +
        lead promoter: no colonies seen
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; RBS-YFP (X/P) Concentration: 18.2 ng/&#956;l
+
    July 20
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; (wrong enzymes used in digestions - plasmid isolation repeated)
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Ligation
+
    Glycerol stocks &amp; plasmid isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLas-TetR: Concentrations: (2:1) 67.4 and (4:1) 68.0
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; Vector: pLac
+
    Digestion
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        (2:1) pLac-TetR
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; Insert: LuxR
+
    Gel Extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-TetR: Concentration: 20.5 ng/μl
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector: pLac-TetR
 +
    </li>
 +
    <li>
 +
        Insert: RBS-YFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Transformation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-TetR-RBS-YFP
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            (0:1) no colonies seen
 +
        </li>
 +
</div>
</div>
 +
<div id="page11" class='notebook_page'>
-
<div id="page6" class='notebook_page'><p>
+
        <li>
-
Transformation
+
            (2:1) 1 colony picked
 +
        </li>
 +
    </ul>
 +
</ul>
 +
<br/>
 +
<p>
 +
    Single and Double Population Projects
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; BBa_ K117002: pLsrA
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; BBa_R0062: pLuxR
+
    July 20
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; BBa_I721001: lead promoter
+
    Repeat Transformation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        lead promoter
 +
    </li>
 +
    <li>
 +
        LasI
 +
    </li>
 +
    <li>
 +
        RBS-YFP
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
Colonies picked
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR: 15 colonies seen: 1 colony picked
+
    July 21
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLsrA: 20 colonies seen: colony picked
+
    Glycerol Stock
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
    <li>
 +
        LuxR
 +
    </li>
 +
    <li>
 +
        LuxI
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pLuxR: 3 colonies seen: 1 colony picked
+
    Colonies picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        RBS-YFP: 1 colony picked
 +
    </li>
 +
 +
</div>
 +
<div id="page12" class='notebook_page'>
 +
 +
    <li>
 +
        LasI: 1 colony picked
 +
    </li>
 +
    <li>
 +
        lead promoter: no colones seen
 +
    </li>
 +
</ul>
<p>
<p>
-
<strong>July 24</strong>
+
    Digestion &amp; Gel Extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS (wrong enzymes used)
 +
    </li>
 +
    <li>
 +
        LuxR: Concentration: 11.1 ng/μl
 +
    </li>
 +
    <li>
 +
        LuxI: Concentration: 7.1 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
 +
</div>
 +
<div id="page13" class='notebook_page'>
 +
<p>
<p>
-
Plasmid Isolation
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: Concentration: 85.9 ng/&#956;l
+
 
 +
 
 +
    July 22
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxI: Concentration:85.7 ng/&#956;l
+
    Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS: Concentration: 75.2 ng/μl
 +
    </li>
 +
    <li>
 +
        LasI: Concentration: 90.1 ng/μl
 +
    </li>
 +
    <li>
 +
        RBS-YFP: Concentration: 76.4 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; LasI: Concentration: 224.1 ng/&#956;l
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Digestion and gel extraction
+
    July 23
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxI (S/P): Concentration: 25.5 ng/&#956;l
+
    DIgestion and gel extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS (X/P) Concentration: not right size to be repeated
 +
    </li>
 +
    <li>
 +
        LasI (E/S) Concentration: 20.3 ng/μl
 +
    </li>
 +
    <li>
 +
        RBS-YFP (X/P) Concentration: 18.2 ng/μl
 +
    </li>
 +
    <li>
 +
        (wrong enzymes used in digestions - plasmid isolation repeated)
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; LasI (S/P): Concentration: 30.7 ng/&#956;l
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector: pLac
 +
    </li>
 +
    <li>
 +
        Insert: LuxR
 +
    </li>
 +
</ul>
 +
<p>
 +
    Transformation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        BBa_ K117002: pLsrA
 +
    </li>
 +
</div>
</div>
 +
<div id="page14" class='notebook_page'>
-
<div id="page7" class='notebook_page'><p>
+
    <li>
-
Ligation
+
        BBa_R0062: pLuxR
 +
    </li>
 +
    <li>
 +
        BBa_I721001: lead promoter
 +
    </li>
 +
</ul>
 +
<p>
 +
    Colonies picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR: 15 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        pLsrA: 20 colonies seen: colony picked
 +
    </li>
 +
    <li>
 +
        pLuxR: 3 colonies seen: 1 colony picked
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; Vector 1: LuxI
+
    July 23
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert 1: RBS-YFP
+
    Glycerol stock and plasmid isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-TetR-RBS-YFP: Concetration: 138.4
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; Vector 2: LasI
+
    Digestion
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-TetR-RBS-YFP (X/P)
 +
    </li>
 +
    <li>
 +
        pTetR-LacI-RBS-GFP (S/P)
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; Insert 2: RBS-RFP
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector: pTetR-LacI-RBS-GFP
 +
    </li>
 +
    <li>
 +
        Insert: pLac-TetR-RBS-YFP
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            2 colonies picked after plating and incubation
 +
        </li>
 +
 +
</div>
 +
<div id="page15" class='notebook_page'>
 +
 +
    </ul>
 +
</ul>
<p>
<p>
-
<strong>July 25</strong>
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Colonies picked
+
    July 24
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxI-RBS-YFP: 100 colonies seen: 1 picked
+
    Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS: Concentration: 85.9 ng/μl
 +
    </li>
 +
    <li>
 +
        LuxI: Concentration:85.7 ng/μl
 +
    </li>
 +
    <li>
 +
        LasI: Concentration: 224.1 ng/μl
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; LasI-RBS-RFP: 50 colonies seen: 1 picked
+
    Digestion and gel extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxI (S/P): Concentration: 25.5 ng/μl
 +
    </li>
 +
    <li>
 +
        LasI (S/P): Concentration: 30.7 ng/μl
 +
    </li>
 +
</ul>
<p>
<p>
-
Plasmid Isolation
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector 1: LuxI
 +
    </li>
 +
    <li>
 +
        Insert 1: RBS-YFP
 +
    </li>
 +
    <li>
 +
        Vector 2: LasI
 +
    </li>
 +
    <li>
 +
        Insert 2: RBS-RFP
 +
    </li>
 +
</ul>
 +
<br/>
 +
 +
</div>
 +
<div id="page16" class='notebook_page'>
 +
<p>
<p>
-
&#9679; pLac-LuxR: Concentration: 107.5
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLsrA: Concentration: 44.3
+
    July 24
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR: Concentration: 35.9
+
 
 +
    Transformation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        BBa_C0079: LasR
 +
    </li>
 +
    <li>
 +
        BBa_B0015: Term-Term
 +
    </li>
 +
    <li>
 +
        BBa_I14016: pLac
 +
    </li>
 +
    <li>
 +
        BBa_C0012: LacI
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
Digestion and gel extraction
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLsrA (S/P): Concentration: 20.6 ng/&#956;l
+
    July 25
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR (S/P): Concentration: 18.7 ng/&#956;l
+
    Colonies picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxI-RBS-YFP: 100 colonies seen: 1 picked
 +
    </li>
 +
    <li>
 +
        LasI-RBS-RFP: 50 colonies seen: 1 picked
 +
    </li>
 +
</ul>
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR: Concentration: 107.5
 +
    </li>
 +
    <li>
 +
        pLsrA: Concentration: 44.3
 +
    </li>
 +
    <li>
 +
        pLuxR: Concentration: 35.9
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion and gel extraction
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLsrA (S/P): Concentration: 20.6 ng/μl
 +
    </li>
 +
</div>
</div>
 +
<div id="page17" class='notebook_page'>
-
<div id="page8" class='notebook_page'><p>
+
    <li>
-
Transformation
+
        pLuxR (S/P): Concentration: 18.7 ng/μl
-
</p>
+
    </li>
 +
</ul>
<p>
<p>
-
&#9679; BBa_R0040: pTetR
+
    Transformation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        BBa_R0040: pTetR
 +
    </li>
 +
    <li>
 +
        BBa_R0079: pLasR
 +
    </li>
 +
    <li>
 +
        BBa_K17036: RBS-CFP
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; BBa_R0079: pLasR
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; BBa_K17036: RBS-CFP
+
    July 25
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>July 26</strong>
+
    Glycerol Stock and plasmid isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-LacI-RBS-GFP-pLac-TetR-RBS-YFP (full toggle switch) from both colines picked
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            Concentration 1: 194.8 ng/μl
 +
        </li>
 +
        <li>
 +
            Concentartion 2: 117.2 ng/μl
 +
        </li>
 +
    </ul>
 +
</ul>
<p>
<p>
-
Colonies picked
+
    Transformation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        BBa_C0079: LasR
 +
    </li>
 +
    <li>
 +
        BBa_I14016: pLas
 +
    </li>
 +
    <li>
 +
        BBa_C0012: LacI
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pTetR: 10 colonies seen: 1 colony picked
+
    Tetracycle Stock: 50 ng/чl
</p>
</p>
 +
<br/>
 +
 +
</div>
 +
<div id="page18" class='notebook_page'>
<p>
<p>
-
&#9679; pLasR: 10 colonies seen: 1 colony picked
+
    IPTG stock: 50 ng/чl
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; RBS-CFP: no colonies seen
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Glycerol stock and plasmid isolation
+
    July 26
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxI-RBS-YFP: Concentration: 70.8 ng/&#956;l
+
    Colonies picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR: 10 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        pLasR: 10 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        RBS-CFP: no colonies seen
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; LasI-RBS-RFP: Concentration: 4.0 (to be repeated) ng/&#956;l
+
    Glycerol stock and plasmid isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxI-RBS-YFP: Concentration: 70.8 ng/μl
 +
    </li>
 +
    <li>
 +
        LasI-RBS-RFP: Concentration: 4.0 (to be repeated) ng/μl
 +
    </li>
 +
</ul>
<p>
<p>
-
Digestion and gel extraction
+
    Digestion and gel extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxI-RBS-YFP (X/P): Concentration: 2.8 (to be repeated)
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; LuxI-RBS-YFP (X/P): Concentration: 2.8 (to be repeated)
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>July 27</strong>
+
    July 26
</p>
</p>
 +
<br/>
<p>
<p>
-
Glycerol stock and plasmid isolation
+
    Transformation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        BBa_C0012: LacI
 +
    </li>
</div>
</div>
 +
<div id="page19" class='notebook_page'>
-
<div id="page9" class='notebook_page'><p>
+
    <li>
-
&#9679; pTetR: Concentration: 42.4 ng/&#956;l
+
        BBa_I732100: LacI (2)
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<p>
 +
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLasR: Concentration: 62.2 ng/&#956;l
+
    July 27
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxI-RBS-YFP: Concentration: 120.9 ng/&#956;l
+
    Glycerol stock and plasmid isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR: Concentration: 42.4 ng/μl
 +
    </li>
 +
    <li>
 +
        pLasR: Concentration: 62.2 ng/μl
 +
    </li>
 +
    <li>
 +
        LuxI-RBS-YFP: Concentration: 120.9 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LasI-RBS-RFP: Concentration: 85 ng/μl
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; LasI-RBS-RFP: Concentration: 85 ng/&#956;l
+
    Digestion and gel extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR (S/P): Concentration: 17.4 ng/μl
 +
    </li>
 +
    <li>
 +
        pLasR (S/P): Concentration: 23.8 ng/μl
 +
    </li>
 +
    <li>
 +
        LuxI-RBS-YFP (X/P): Concentration: 13.8 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LasI-RBS-RFP (X/P): Concentration: 8.9 ng/μl
 +
    </li>
 +
</ul>
<p>
<p>
-
Digestion and gel extraction
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector 1: pLsrA
 +
    </li>
 +
    <li>
 +
        Insert 1:LuxI-RBS-YFP
 +
    </li>
 +
 +
</div>
 +
<div id="page20" class='notebook_page'>
 +
 +
    <li>
 +
        Vector 2: pLuxR
 +
    </li>
 +
    <li>
 +
        Insert 2:LasI-RBS-RFP
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pTetR (S/P): Concentration: 17.4 ng/&#956;l
+
    Transformation:
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLsrA-LuxI-RBS-YFP
 +
    </li>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        BBa_B0422: RBS-CFP
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLasR (S/P): Concentration: 23.8 ng/&#956;l
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxI-RBS-YFP (X/P): Concentration: 13.8 ng/&#956;l
+
    July 27
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LasI-RBS-RFP (X/P): Concentration: 8.9 ng/&#956;l
+
    Colonies picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LacI: 8 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        LacI (2): 2 colonies seen: 1 colony picked
 +
    </li>
 +
</ul>
<p>
<p>
-
Ligation
+
    Plasmid isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LasR: Concentration: 95.8 ng/μl
 +
    </li>
 +
    <li>
 +
        Term-Term: Concentration: 120.1 ng/μl
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; Vector 1: pLsrA
+
    Toggle switch test
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        IPTG innoculation
 +
    </li>
 +
</div>
 +
<div id="page21" class='notebook_page'>
 +
 +
    <li>
 +
        TET innoculation
 +
    </li>
 +
    <li>
 +
        No innoculation
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; Insert 1:LuxI-RBS-YFP
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector 2: pLuxR
+
    July 28
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert 2:LasI-RBS-RFP
+
    Colonies picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLsrA-LuxI-RBS-YFP: 20 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP: 2 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        RBS-CFP: 10 colonies seen: 1 colony picked
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
<p>
<p>
-
Transformation:
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLsrA-LuxI-RBS-YFP
+
    July 28
</p>
</p>
 +
<br/>
 +
<p>
 +
    Glycerol stock and plasmid isolation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LacI: Concentration: 73.9 ng/μl
 +
    </li>
 +
    <li>
 +
        LacI (2): Concentration: 109.8 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
</div>
</div>
 +
<div id="page22" class='notebook_page'>
-
<div id="page10" class='notebook_page'><p>
+
<p>
-
&#9679; pLuxR-LasI-RBS-RFP
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; BBa_B0422: RBS-CFP
+
    July 29
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>July 28</strong>
+
    Glycerol Stocks
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLsrA-LuxI-RBS-YFP: Concentration: 201.6 ng/μl
 +
    </li>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP: Concentration: 127.1 ng/μl
 +
    </li>
 +
    <li>
 +
        RBS-CFP: Concentration: 84.0 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
Colonies picked
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLsrA-LuxI-RBS-YFP: 20 colonies seen: 1 colony picked
+
    July 30
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-LasI-RBS-RFP: 2 colonies seen: 1 colony picked
+
    Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLsrA-LuxI-RBS-YFP: Concentration: 201.6 ng/μl
 +
    </li>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP: Concentration: 127.1 ng/μl
 +
    </li>
 +
    <li>
 +
        RBS-CFP: Concentration: 84.0 ng/μl
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion and gel extraction
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS (X/P): Gel extraction failed
 +
    </li>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP (X/P): Gel extraction failed
 +
    </li>
 +
    <li>
 +
        RBS-CFP (X/P): Concentration: 10.4 ng/μl
 +
    </li>
 +
    <li>
 +
        pLac-LuxR (S/P): Concentration: 21.0 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; RBS-CFP: 10 colonies seen: 1 colony picked
+
    Innoculation
</p>
</p>
 +
<br/>
</div>
</div>
-
<div id="page11" class='notebook_page'><p>
+
<div id="page23" class='notebook_page'>
-
<strong>July 29</strong>
+
 
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP
 +
    </li>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation and transformation:
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector: pLasR
 +
    </li>
 +
    <li>
 +
        Insert: RBS-CFP
 +
    </li>
 +
    <li>
 +
        2 colonies shown: 1 colony picked
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
Glycerol Stocks
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLsrA-LuxI-RBS-YFP: Concentration: 201.6 ng/&#956;l
+
    July 30
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-LasI-RBS-RFP: Concentration: 127.1 ng/&#956;l
+
    Experiment 7
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        150 ng/чl Tet
 +
    </li>
 +
    <li>
 +
        200 ng/чl Tet
 +
    </li>
 +
    <li>
 +
        300 ng/чl Tet
 +
    </li>
 +
    <li>
 +
        1mM IPTG
 +
    </li>
 +
    <li>
 +
        No Induction
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; RBS-CFP: Concentration: 84.0 ng/&#956;l
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        RBS-GFP
 +
    </li>
 +
</div>
 +
<div id="page24" class='notebook_page'>
 +
 +
</ul>
 +
<br/>
<p>
<p>
-
<strong>July 30</strong>
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Plasmid Isolation
+
    July 31
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLsrA-LuxI-RBS-YFP: Concentration: 201.6 ng/&#956;l
+
    Plasmid isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS: Concentration: 78.1 ng/μl
 +
    </li>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP: Concentration: 48.7 ng/μl
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pLuxR-LasI-RBS-RFP: Concentration: 127.1 ng/&#956;l
+
    Digestion:
-
</p>
+
-
</div>
+
-
 
+
-
<div id="page12" class='notebook_page'><p>
+
-
&#9679; RBS-CFP: Concentration: 84.0 ng/&#956;l
+
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS: (X/P); 516 bp
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLuxR+LasI-RBS-RFP: Digested at XbaI and PstI; 1421 bp
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
Digestion and gel extraction
+
    Test Digestion:
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS: digested at EcoRI and PstI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLuxR-lasI-RBS-RFP: digested at EcoRI and PstI
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; LuxS (X/P): Gel extraction failed
+
    Colonies:
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLuxR-lasI-RBS-RFP: 1 additional colony picked
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS: 1 additional colony picked
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
</div>
 +
<div id="page25" class='notebook_page'>
<p>
<p>
-
&#9679; pLuxR-LasI-RBS-RFP (X/P): Gel extraction failed
+
 
 +
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; RBS-CFP (X/P): Concentration: 10.4 ng/&#956;l
+
    July 31
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR (S/P): Concentration: 21.0 ng/&#956;l
+
    Plasmid Isolation:
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        GFP: 137.5 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
Innoculation
+
    August 1, 2012:
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-LasI-RBS-RFP
+
    Plasmid Isolation:
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP: 61.2 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLasR-RBS-CFP: 49.2 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; LuxS
+
    Digestion:
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        GFP: Digested XbaI and PstI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LacI: Digested XbaI and PstI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP: Digested XbaI and PstI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLusR-RBS-CFP: Digested XbaI and PstI
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
Ligation and transformation:
+
    Gel Exraction:
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        GFP: 8.5 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LacI: 12.7 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
 +
 +
</div>
 +
<div id="page26" class='notebook_page'>
<p>
<p>
-
&#9679; Vector: pLasR
+
    Colony:
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLasR-RBS-CFP: 1 additional colony picked
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; Insert: RBS-CFP
+
    Transformation:
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLuxR-lasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        BBa_J2309: pTetR-luxI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; 2 colonies shown: 1 colony picked
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>July 31</strong>
+
    August 1
</p>
</p>
 +
<br/>
<p>
<p>
-
Plasmid isolation
+
    Ligation:
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector: pTet; Insert: GFP
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector: pTet; Insert: LacI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector: pLac; Insert: GFP
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: Concentration: 78.1 ng/&#956;l
+
    Transformation:
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTet-GFP
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTet-LacI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-GFP
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
</div>
</div>
 +
<div id="page27" class='notebook_page'>
-
<div id="page13" class='notebook_page'><p>
+
<p>
-
&#9679; pLuxR-LasI-RBS-RFP: Concentration: 48.7 ng/&#956;l
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Digestion:
+
    August 2, 2012:
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
-
LuxS: (X/P); 516 bp
+
-
</li>
+
-
<li>
+
-
pLuxR+LasI-RBS-RFP: Digested at XbaI and PstI; 1421 bp
+
-
</li>
+
-
</ul>
+
<p>
<p>
-
Test Digestion:
+
    Colonies:
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<ul>
-
LuxS: digested at EcoRI and PstI
+
    <li>
-
</li>
+
        pLuxR-LasI-RBS-RFP: about 5 colonies; 1 picked
-
<li>
+
    </li>
-
pLuxR-lasI-RBS-RFP: digested at EcoRI and PstI
+
</ul>
-
</li>
+
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-LuxI: about 2 colonies; 1 picked
 +
    </li>
</ul>
</ul>
 +
<br/>
<p>
<p>
-
Colonies:
+
    Plasmid Isolation:
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<ul>
-
pLuxR-lasI-RBS-RFP: 1 additional colony picked
+
    <li>
-
</li>
+
        LuxS: Concentration – 40.5 ng/μl
-
<li>
+
    </li>
-
LuxS: 1 additional colony picked
+
-
</li>
+
</ul>
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLasR-CFP: 95.8 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
<strong>August 1</strong>
+
    Digestion:
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LasI: Digested at XbaI and PstI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LasR: Digested at XbaI and PstI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS: Digested at XbaI and PstI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLasR-RBS-CFP: Digested at XbaI and PstI
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
Plasmid Isolation:
+
    Gel Extraction:
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<ul>
-
pLuxR-LasI-RBS-RFP: 61.2 ng/&#956;l
+
    <li>
-
</li>
+
        LasI: Concentration – 4.1 ng/μl
-
<li>
+
    </li>
-
pLasR-RBS-CFP: 49.2 ng/&#956;l
+
-
</li>
+
</ul>
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LasR: Concentration – 5.3 ng/μl
 +
    </li>
</div>
</div>
-
 
+
<div id="page28" class='notebook_page'>
-
<div id="page14" class='notebook_page'><p>
+
</ul>
-
Digestion:
+
<br/>
 +
<ul>
 +
    <li>
 +
        pLasR-RBS-CFP – 13.4 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
 +
<p>
 +
    Ligation:
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<ul>
-
GFP: Digested XbaI and PstI
+
    <li>
-
</li>
+
        Vector: pLac
-
<li>
+
    </li>
-
LacI: Digested XbaI and PstI
+
-
</li>
+
-
<li>
+
-
pLuxR-LasI-RBS-RFP: Digested XbaI and PstI
+
-
</li>
+
-
<li>
+
-
pLusR-RBS-CFP: Digested XbaI and PstI
+
-
</li>
+
</ul>
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Insert: LasI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Transformation 2:1 – 3 colonies; 1 picked
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
Gel Extraction:
+
    Single and Double Population Projects
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<p>
-
GFP: 8.5 ng/&#956;l
+
    August 2
-
</li>
+
</p>
-
<li>
+
<br/>
-
LacI: 12.7 ng/&#956;l
+
<p>
-
</li>
+
    Colonies:
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTet-GFP: 7 colonies; 1 picked
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTet-LacI: 20 colonies; 1 picked
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-GFP: 20 colonies; 1 picked
 +
    </li>
</ul>
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
Colony:
+
    Single and Double Population Projects
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<p>
-
pLasR-RBS-CFP: 1 additional colony picked
+
    August 3, 2012:
-
</li>
+
</p>
 +
<br/>
 +
<p>
 +
    Glycerol Stocks/Miniprep:
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTet-GFP: Concentration-142.2 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTet-LacI: Concentration-67.2 ng/μl
 +
    </li>
 +
</div>
 +
<div id="page29" class='notebook_page'>
 +
 
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-GFP: Concentration-50.5 ng/μl
 +
    </li>
</ul>
</ul>
 +
<br/>
<p>
<p>
-
Transformation:
+
    Digestion:
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<ul>
-
pLuxR-lasI-RBS-RFP
+
    <li>
-
</li>
+
        pTet-GFP: Digested at SpeI and PstI
-
<li>
+
    </li>
-
BBa_J2309: pTetR-luxI
+
-
</li>
+
</ul>
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTet-LacI: Digested at SpeI and PstI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-GFP: Digested at XbaI and PstI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-TetR: Digested at XbaI and PstI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
<p>
<p>
-
<strong>August 2</strong>
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Colonies:
+
    August 3
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<p>
-
pLuxR-LasI-RBS-RFP: about 5 colonies; 1 picked
+
    Ligation:
-
</li>
+
</p>
-
<li>
+
<br/>
-
pTetR-LuxI: about 2 colonies; 1 picked
+
<ul>
-
</li>
+
    <li>
 +
        Vector: pTetR
 +
    </li>
</ul>
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Insert: LasR
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Transformation 2:1 – 1 colony; 1 picked
 +
    </li>
 +
</ul>
 +
<br/>
 +
<p>
 +
    Ligation:
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector: pLasR
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Insert: RBS-CFP
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Transformation: 2:1
 +
    </li>
 +
</ul>
 +
<br/>
</div>
</div>
 +
<div id="page30" class='notebook_page'>
-
<div id="page15" class='notebook_page'><p>
+
<p>
-
Plasmid Isolation:
+
    Glycerol Stock:
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<ul>
-
LuxS: Concentration - 40.5 ng/&#956;l
+
    <li>
-
</li>
+
        pLuxR-LasI-RBS-RFP: Concentration – 45.1 ng/μl
-
<li>
+
    </li>
-
pLasR-CFP: 95.8 ng/&#956;l
+
-
</li>
+
</ul>
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-LuxI: Concentration – 105.1 ng/μl
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
Digestion:
+
    Digestion:
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<ul>
-
LasI: Digested at XbaI and PstI
+
    <li>
-
</li>
+
        pLuxR-LasI-RBS-RFP: Digested at XbaI and PstI
-
<li>
+
    </li>
-
LasR: Digested at XbaI and PstI
+
-
</li>
+
-
<li>
+
-
LuxS: Digested at XbaI and PstI
+
-
</li>
+
-
<li>
+
-
pLasR-RBS-CFP: Digested at XbaI and PstI
+
-
</li>
+
</ul>
</ul>
 +
<br/>
<p>
<p>
-
Gel Extraction:
+
    Ligation:
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<ul>
-
LasI: Concentration - 4.1 ng/&#956;l
+
    <li>
-
</li>
+
        Vector: pLac-luxR
-
<li>
+
    </li>
-
LasR: Concentration - 5.3 ng/&#956;l
+
    <li>
-
</li>
+
        Insert: pLuxR-LasI-RBS-RFP
-
<li>
+
    </li>
-
pLasR-RBS-CFP - 13.4 ng/&#956;l
+
-
</li>
+
</ul>
</ul>
<p>
<p>
-
Ligation:
+
    Transformation
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<ul>
-
Vector: pLac
+
    <li>
-
</li>
+
        BBa_K091109 (2010) LuxS
-
<li>
+
    </li>
-
Insert: LasI
+
    <li>
-
</li>
+
        BBa_K091109 (2011) LuxS
-
<li>
+
    </li>
-
Transformation 2:1 - 3 colonies; 1 picked
+
-
</li>
+
</ul>
</ul>
-
</div>
+
<br/>
-
<div id="page16" class='notebook_page'><p>
+
<p>
-
<strong>August 3</strong>
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Ligation:
+
    August 4
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<p>
-
Vector: pTetR
+
    Colonies picked
-
</li>
+
</p>
-
<li>
+
<br/>
-
Insert: LasR
+
<ul>
-
</li>
+
    <li>
-
<li>
+
        BBa_K091109 (2010) LuxS: 70 colonies seen: 1 picked
-
Transformation 2:1 - 1 colony; 1 picked
+
    </li>
-
</li>
+
    <li>
 +
        BBa_K091109 (2011) LuxS: 100 colonies seen: 1 picked
 +
    </li>
</ul>
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
 +
<br/>
 +
 +
</div>
 +
<div id="page31" class='notebook_page'>
 +
<p>
<p>
-
Ligation:
+
    Triple Population and AND Gate Projects
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<p>
-
Vector: pLasR
+
    August 5
-
</li>
+
</p>
-
<li>
+
<br/>
-
Insert: RBS-CFP
+
<p>
-
</li>
+
    Glycerol Stock
-
<li>
+
</p>
-
Transformation: 2:1
+
<br/>
-
</li>
+
<ul>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
    <li>
 +
        pTetR-RBS-CFP
 +
    </li>
</ul>
</ul>
<p>
<p>
-
Glycerol Stock:
+
    Transformation (repeated)
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<ul>
-
pLuxR-LasI-RBS-RFP: Concentration - 45.1 ng/&#956;l
+
    <li>
-
</li>
+
        pLasR-RBS-CFP
-
<li>
+
    </li>
-
pTetR-LuxI: Concentration - 105.1 ng/&#956;l
+
    <li>
-
</li>
+
        <br/>
 +
    </li>
</ul>
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
Digestion:
+
    Triple Population and AND Gate Projects
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<p>
-
pLuxR-LasI-RBS-RFP: Digested at XbaI and PstI
+
    August 6
-
</li>
+
</p>
 +
<br/>
 +
<p>
 +
    Colonies picked
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLasR-RBS-CFP: 10 colonies seen: 1 colony picked
 +
    </li>
</ul>
</ul>
 +
<br/>
<p>
<p>
-
Ligation:
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; Vector: pLac-luxR
+
    Plasmid isolation
</p>
</p>
-
</div>
+
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS: Concentration: 57.6
 +
    </li>
-
<div id="page17" class='notebook_page'><p>
+
</div>
-
&#9679; Insert: pLuxR-LasI-RBS-RFP
+
<div id="page32" class='notebook_page'>
-
</p>
+
    <li>
 +
        LuxS: Concentration: 76.4
 +
    </li>
 +
    <li>
 +
        pTetR-RBS-CFP: Concentration: 122.3
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
Transformation
+
    Digestion and gel extraction (improperly prepared for gel)
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS: Concentration: N/A
 +
    </li>
 +
    <li>
 +
        LuxS: Concentration: N/A
 +
    </li>
 +
    <li>
 +
        pTetR-RBS-CFP: Concentration: 21.9
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; BBa_K091109 (2010) LuxS
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
    <li>
 +
        pTetR-RBS-CFP
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; BBa_K091109 (2011) LuxS
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>August 4</strong>
+
    August 6, 2012:
</p>
</p>
 +
<br/>
<p>
<p>
-
Colonies picked
+
    PCR:
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; BBa_K091109 (2010) LuxS: 70 colonies seen: 1 picked
+
    RFP-LVA created
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; BBa_K091109 (2011) LuxS: 100 colonies seen: 1 picked
+
    Experiments:
</p>
</p>
 +
<br/>
 +
<p>
 +
    Test mechanism shows TetR and LacI are medium strength repressors.
 +
</div>
</div>
 +
<div id="page33" class='notebook_page'>
-
<div id="page18" class='notebook_page'><p>
 
-
<strong>August 5</strong>
 
</p>
</p>
-
<p>
+
<br/>
-
Glycerol Stock
+
<p dir="ltr">
 +
    Toggle Experiment fails to show significant changes in fluorescence protein after 2 or 6 hours.
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS
+
    August 7
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-RBS-CFP
+
    Glycerol Stock
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLasR-RBS-CFP
 +
    </li>
 +
</ul>
<p>
<p>
-
Transformation (repeated)
+
    Plasmid isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS: Concentration: 120.3
 +
    </li>
 +
    <li>
 +
        LuxS: Concentration:101.7
 +
    </li>
 +
    <li>
 +
        pTetR-LasR: Concentration: 65.3
 +
    </li>
 +
    <li>
 +
        pLasR-RBS-CFP: Concentration: 77.1
 +
    </li>
 +
    <li>
 +
        pLac-luxR-pLuxR-LasI-RBS-RFP: Concentration: 103.6
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pLasR-RBS-CFP
+
    Digestion and gel extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        LuxS (X/P): Concentration: 1.5
 +
    </li>
 +
    <li>
 +
        LuxS (X/P): Concentration: 1.1
 +
    </li>
 +
    <li>
 +
        pTetR-LasR (S/P): Concentration: 20.4
 +
    </li>
 +
</div>
 +
<div id="page34" class='notebook_page'>
 +
 +
    <li>
 +
        pLasR-RBS-CFP (X/P): Concentration: 13.8
 +
    </li>
 +
</ul>
<p>
<p>
-
<strong>August 6</strong>
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector 1: pTetR-lasR
 +
    </li>
 +
    <li>
 +
        Insert 1: pLasR-RBS-CFP
 +
    </li>
 +
    <li>
 +
        Vector 2: pyear
 +
    </li>
 +
    <li>
 +
        Insert 2: LuxS
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
Colonies picked
+
    Transformation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pSBIC3 chlora backbone
 +
    </li>
 +
    <li>
 +
        pLac-luxR-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLasR-RBS-CFP: 10 colonies seen: 1 colony picked
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Innoculation
+
    August 7, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LuxI
+
    Glycerol Stock/Miniprep
</p>
</p>
 +
<br/>
<p>
<p>
-
Plasmid isolation
+
    pTet-LacI
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: Concentration: 57.6
+
    pLac-GFP
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: Concentration: 76.4
+
    Digestion &amp; Gel Extraction
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-RBS-CFP: Concentration: 122.3
+
    pTet-LacI: XbaI and PstI
</p>
</p>
 +
<br/>
</div>
</div>
 +
<div id="page35" class='notebook_page'>
-
<div id="page19" class='notebook_page'><p>
+
<p>
-
Digestion and gel extraction (improperly prepared for gel)
+
    pLac-GFP: SpeI and PstI
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: Concentration: N/A
+
    Ligation/Transformation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: Concentration: N/A
+
    Vector: pLac-GFP
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-RBS-CFP: Concentration: 21.9
+
    Insert: pTet-LacI
</p>
</p>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
Innoculation
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS
+
    August 8
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS
+
    Colonies picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        chlora bb: 100 colonies seen: 1 picked
 +
    </li>
 +
    <li>
 +
        pLac-luxR-pLuxR-LasI-RBS-RFP: 300 colonies seen: 1 picked
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-pLasR-RBS-CFP: 1 colony seen: 1 picked
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pTetR-RBS-CFP
+
    Repeat transformations from ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pyear-LuxS
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-pLasR-RBS-CFP
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
<strong>August 7</strong>
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Glycerol Stock
+
    August 8, 2012
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLasR-RBS-CFP
+
    Transformations:
 +
 
 +
</div>
 +
<div id="page36" class='notebook_page'>
</p>
</p>
 +
<br/>
<p>
<p>
-
Plasmid isolation
+
    Negative control had ~20 colonies
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: Concentration: 120.3
+
    2:1 sample had ~ 40 colonies, we picked 2
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: Concentration:101.7
+
    Positive had too many colonies to count
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LasR: Concentration: 65.3
+
    PCR Reaction (Gradient)
</p>
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    -55, 60, and 65 degrees
 +
</p>
 +
<p dir="ltr">
 +
    -55 degrees found to be most ideal
 +
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLasR-RBS-CFP: Concentration: 77.1
+
    Gel Extraction (of PCR)
</p>
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    Poor results, no DNA extracted
 +
</p>
 +
<br/>
 +
<p>
 +
    Plate test for toggle switch:
 +
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    1 tube inoculated with IPTG 10 microliters of .2mM solution
 +
</p>
 +
<p dir="ltr">
 +
    1 tube inoculated with ATC 10 microliters of 20ng/microliter
 +
</p>
 +
<br/>
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
<br/>
 +
<p>
 +
    August 9
 +
</p>
 +
<br/>
 +
<p>
 +
    Glycerol stocks
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        chlora backbone
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-pLuxR-LasI-RBS-RFP
 +
    </li>
</div>
</div>
 +
<div id="page37" class='notebook_page'>
-
<div id="page20" class='notebook_page'><p>
+
    <li>
-
&#9679; pLac-luxR-pLuxR-LasI-RBS-RFP: Concentration: 103.6
+
        pTetR-LasR-pLasR-RBS-CFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-Las-I
 +
    </li>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
</ul>
<p>
<p>
-
Digestion and gel extraction
+
    Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        chlora backbone
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
    <li>
 +
        pTetR-LasR-pLasR-RBS-CFP
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; LuxS (X/P): Concentration: 1.5
+
    Digestion
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Term-Term (S/P)
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        chlora backbone (E/P)
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-pLuxR-LasI-RBS-RFP (E/P)
 +
    </li>
 +
    <li>
 +
        pTetR-LasR-pLasR-RBS-CFP (E/P)
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; LuxS (X/P): Concentration: 1.1
+
    Gel Extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Term-Term: Concentration: 44.3
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        chlora backbone: Concentration: 332.9
 +
    </li>
 +
</div>
 +
<div id="page38" class='notebook_page'>
 +
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-pLuxR-LasI-RBS-RFP: bad results
 +
    </li>
 +
    <li>
 +
        pTetR-LasR-pLasR-RBS-CFP: bad results
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pTetR-LasR (S/P): Concentration: 20.4
+
    Innoculation: pLac-LuxR-pLuxR-LasI-RBS-RFP
</p>
</p>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLasR-RBS-CFP (X/P): Concentration: 13.8
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Ligation
+
    August 9, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector 1: pTetR-lasR
+
    Glycerol stock/miniprep:
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert 1: pLasR-RBS-CFP
+
    pLac-GFP-ST-pTet-LacI
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; Vector 2: pyear
+
    PCR Reaction
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert 2: LuxS
+
    RFP-LVA
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
Transformation
+
    Experiment: Repressor Test Mechanism (LacI)
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pSBIC3 chlora backbone
+
    1 - Induced 10microliters of 10mM IPTG
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-luxR-pLuxR-LasI-RBS-RFP
+
    1 - No induction
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
<strong>August 8</strong>
+
    Digestion and Gel Extraction:
</p>
</p>
 +
<br/>
<p>
<p>
-
Colonies picked
+
    RFP-LVA: XbaI and PstI
</p>
</p>
 +
<br/>
 +
<p>
 +
    Ampicillin Backbone: SpeI and PstI
 +
</p>
 +
<br/>
 +
<br/>
 +
<p>
 +
</div>
</div>
 +
<div id="page39" class='notebook_page'>
-
<div id="page21" class='notebook_page'><p>
+
    Triple Population and AND Gate Projects
-
&#9679; chlora bb: 100 colonies seen: 1 picked
+
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-luxR-pLuxR-LasI-RBS-RFP: 300 colonies seen: 1 picked
+
    August 10
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-lasR-pLasR-RBS-CFP: 1 colony seen: 1 picked
+
    Glycerol Stock
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
<p>
<p>
-
Repeat transformations from ligation
+
    Plasmid isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-pLuxR-LasI-RBS-RFP: Concentration: 125.1
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pyear-LuxS
+
    Digestion
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-pLuxR-LasI-RBS-RFP (E/P)
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pTetR-lasR-pLasR-RBS-CFP
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector1: Term-Term
 +
    </li>
 +
    <li>
 +
        Insert1: pLasR-RBS-CFP
 +
    </li>
 +
    <li>
 +
        Vector2: Term-Term
 +
    </li>
 +
    <li>
 +
        Insert2: pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
<p>
<p>
-
<strong>August 9</strong>
+
    Transformation:
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Term-Term-pLasR-RBS-CFP
 +
    </li>
 +
    <li>
 +
        Term-Term-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</div>
 +
<div id="page40" class='notebook_page'>
 +
 +
</ul>
 +
<br/>
<p>
<p>
-
Glycerol stocks
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; chlora backbone
+
    August 10, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-pLuxR-LasI-RBS-RFP
+
    Ligation/Transformation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LasR-pLasR-RBS-CFP
+
    RFP-LVA into Ampicillin backbone
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
Innoculation
+
    Inoculations:
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-Las-I
+
    Toggle
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LuxI
+
    pLas
</p>
</p>
 +
<br/>
<p>
<p>
-
Plasmid Isolation
+
    LacI
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; chlora backbone
+
    LuxR
</p>
</p>
 +
<br/>
 +
<p>
 +
    pTet
 +
</p>
 +
<br/>
 +
<p>
 +
    pLux
 +
</p>
 +
<br/>
 +
<p>
 +
    LuxI
 +
</p>
 +
<br/>
 +
<br/>
 +
<p>
 +
    August 11, 2012
 +
</p>
 +
<br/>
 +
<p>
 +
    Plasmid isolation
 +
</p>
 +
<br/>
 +
<p>
 +
    Toggle
 +
</p>
 +
<br/>
 +
<p>
 +
    pLas
</div>
</div>
 +
<div id="page41" class='notebook_page'>
-
<div id="page22" class='notebook_page'><p>
 
-
&#9679; pLac-LuxR-pLuxR-LasI-RBS-RFP
 
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LasR-pLasR-RBS-CFP
+
    LacI
</p>
</p>
 +
<br/>
<p>
<p>
-
Digestion
+
    LuxR
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Term-Term (S/P)
+
    pTet
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; chlora backbone (E/P)
+
    pLux
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-pLuxR-LasI-RBS-RFP (E/P)
+
    LuxI
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LasR-pLasR-RBS-CFP (E/P)
+
    Digestion/Extraction
</p>
</p>
 +
<br/>
<p>
<p>
-
Gel Extraction
+
    LuxR: X and P
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Term-Term: Concentration: 44.3
+
    pLux: S and P
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; chlora backbone: Concentration: 332.9
+
    LuxI: S and P
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-pLuxR-LasI-RBS-RFP: bad results
+
    LasR: X and P
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LasR-pLasR-RBS-CFP: bad results
+
    Double Terminators (DT): X and P
</p>
</p>
 +
<br/>
<p>
<p>
-
Innoculation:
+
    LasI: X and P
</p>
</p>
-
<ul type="disc">
+
<br/>
-
<li>
+
<br/>
-
pLac-LuxR-pLuxR-LasI-RBS-RFP
+
<br/>
-
</li>
+
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
<br/>
 +
<p>
 +
    August 11
 +
</p>
 +
<br/>
 +
<p>
 +
    Colonies picked
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Term-Term-pLasR-RBS-CFP: 200 colonies seen: 1 picked
 +
    </li>
 +
    <li>
 +
 
 +
        Term-Term-pLuxR-LasI-RBS-RFP: 100 colonies seen: 1 picked
 +
    </li>
</ul>
</ul>
 +
<br/>
</div>
</div>
 +
<div id="page42" class='notebook_page'>
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
<br/>
 +
<p>
 +
    August 12
 +
</p>
 +
<br/>
 +
<p>
 +
    Glycerol Stock and Plasmid isolation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Term-Term-pLasR-RBS-CFP: Concentration: 219.1
 +
    </li>
 +
    <li>
 +
        Term-Term-pLuxR-LasI-RBS-RFP: Concentration: 107.5
 +
    </li>
 +
</ul>
 +
<p>
 +
    Transformation repeat
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pyear-LuxS
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Term-Term-pLasR-RBS-CFP (X/P)
 +
    </li>
 +
    <li>
 +
        Term-Term-pLuxR-LasI-RBS-RFP (X/P)
 +
    </li>
 +
</ul>
 +
<p>
 +
    Gel Extraction
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Term-Term-pLasR-RBS-CFP: Concentration: 10.2
 +
    </li>
 +
    <li>
 +
        Term-Term-pLuxR-LasI-RBS-RFP: Concentration: 15.0
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector1: pTetR-LasR
 +
    </li>
 +
    <li>
 +
        Insert1: Term-Term-pLasR-RBS-CFP
 +
    </li>
 +
</div>
 +
<div id="page43" class='notebook_page'>
-
<div id="page23" class='notebook_page'><p>
+
    <li>
-
<strong>August 10</strong>
+
        Vector2: pLac-LuxR
 +
    </li>
 +
    <li>
 +
        Insert 2: Term-Term-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
<br/>
 +
<p>
 +
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Glycerol Stock
+
    August 13
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-pLuxR-LasI-RBS-RFP
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LasI
 +
    </li>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
Plasmid isolation
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-pLuxR-LasI-RBS-RFP: Concentration: 125.1
+
    August 13, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
Digestion
+
    Miniprep/Glycerol Stock
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-pLuxR-LasI-RBS-RFP (E/P)
+
    RFP-LVA
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
Ligation
+
    Digestions/Extraction
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector1: Term-Term
+
    pLas: S and P
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert1: pLasR-RBS-CFP
+
    LacI: X and P
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector2: Term-Term
+
    RFP-LVA: E and P
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; Insert2: pLuxR-LasI-RBS-RFP
+
    Inoculations
 +
 
 +
</div>
 +
<div id="page43" class='notebook_page'>
 +
 
</p>
</p>
 +
<br/>
<p>
<p>
-
Transformation:
+
    Chlorabackbone
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Term-Term-pLasR-RBS-CFP
+
    RFP-LVA
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Term-Term-pLuxR-LasI-RBS-RFP
+
    DT
</p>
</p>
 +
<br/>
 +
<br/>
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
<br/>
 +
<p>
 +
    August 14
 +
</p>
 +
<br/>
 +
<p>
 +
    Glycerol Stock and Plasmid isolation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-LasR-Term-Term-pLasR-RBS-CFP: Concentration: 95.4
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP: Concentration: 108.3
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-LasR-Term-Term-pLasR-RBS-CFP: bad results
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP: bad results
 +
    </li>
 +
</ul>
 +
<br/>
 +
<p>
 +
    Parts Recieved
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        BBa_K091159
 +
    </li>
 +
    <li>
 +
        BBa_K091002
 +
    </li>
 +
    <li>
 +
        BBa_I13453
 +
    </li>
 +
    <li>
 +
        BBa_B0034
 +
    </li>
</div>
</div>
 +
<div id="page44" class='notebook_page'>
-
<div id="page24" class='notebook_page'><p>
+
</ul>
-
<strong>August 11</strong>
+
<p>
 +
    Streaking
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        BBa_K091159 pBad-LsrA
 +
    </li>
 +
</ul>
<p>
<p>
-
Colonies picked
+
    Ligation repeat
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector1: pTetR-LasR
 +
    </li>
 +
    <li>
 +
        Insert1: Term-Term-pLasR-RBS-CFP
 +
    </li>
 +
    <li>
 +
        Vector2: pLac-LuxR
 +
    </li>
 +
    <li>
 +
        Insert 2: Term-Term-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; Term-Term-pLasR-RBS-CFP: 200 colonies seen: 1 picked
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Term-Term-pLuxR-LasI-RBS-RFP: 100 colonies seen: 1 picked
+
    August 14, 2012
</p>
</p>
-
</div>
+
<br/>
 +
<p>
 +
    Miniprep/Glycerol Stock
 +
</p>
 +
<br/>
 +
<p>
 +
    Chlorabackbone
 +
</p>
 +
<br/>
 +
<p>
 +
    DT
 +
</p>
 +
<br/>
 +
<br/>
 +
<p>
 +
    Digestion/Extraction:
 +
</p>
 +
<br/>
 +
<p>
 +
    DT: X and P
 +
</p>
 +
<br/>
 +
<p>
 +
    Chlorabackbone: E and P
 +
</p>
 +
<br/>
 +
<p>
 +
    Toggle: E and P
 +
</div>
 +
<div id="page45" class='notebook_page'>
-
<div id="page25" class='notebook_page'><p>
 
-
<strong>August 12</strong>
 
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
Glycerol Stock and Plasmid isolation
+
    Ligation/Transformation:
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Term-Term-pLasR-RBS-CFP: Concentration: 219.1
+
    DT into LuxR
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Term-Term-pLuxR-LasI-RBS-RFP: Concentration: 107.5
+
    TetR into pLux
</p>
</p>
 +
<br/>
<p>
<p>
-
Transformation repeat
+
    LasI into pTet
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pyear-LuxS
+
    LasR into DT
</p>
</p>
 +
<br/>
<p>
<p>
-
Digestion
+
    pLas into LacI
</p>
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    pLac into LuxI
 +
</p>
 +
<br/>
<p>
<p>
-
&#9679; Term-Term-pLasR-RBS-CFP (X/P)
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Term-Term-pLuxR-LasI-RBS-RFP (X/P)
+
    August 15
</p>
</p>
 +
<br/>
<p>
<p>
-
Gel Extraction
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
    <li>
 +
        pLac-LasI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-LasR-Term-Term-pLasR-RBS-CFP
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; Term-Term-pLasR-RBS-CFP: Concentration: 10.2
+
    Colonies picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pBad-LsrA: 30 colonies seen: 1 picked
 +
    </li>
 +
</ul>
 +
 +
</div>
 +
<div id="page46" class='notebook_page'>
 +
<p>
<p>
-
&#9679; Term-Term-pLuxR-LasI-RBS-RFP: Concentration: 15.0
+
    Testing
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Pop2 (w/o TT) / pTetR-LuxI (48 hours of incubation)
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            100/0 no results
 +
        </li>
 +
        <li>
 +
            100/100 no results
 +
        </li>
 +
        <li>
 +
            100/200 slight RFP detected no increase of RFP from 100/0
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        Pop3 (w/o TT) / pLac-LasI (48 hours of incubation)
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            100/0 no results
 +
        </li>
 +
        <li>
 +
            100/100 no results
 +
        </li>
 +
        <li>
 +
            100/200 slight RFP detected no increase of CFP from 100/0
 +
        </li>
 +
    </ul>
 +
</ul>
<p>
<p>
-
Ligation
+
    Streaking
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector1: pTetR-LasR
+
    BBa_K091109
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        SP3011&gt;1D LuxS
 +
    </li>
 +
    <li>
 +
        SP4011&gt;1D LuxS
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; Insert1: Term-Term-pLasR-RBS-CFP
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector2: pLac-LuxR
+
    August 15, 2012
</p>
</p>
 +
<br/>
 +
<p>
 +
    Transformation
 +
</p>
 +
<br/>
</div>
</div>
-
 
+
<div id="page47" class='notebook_page'>
-
<div id="page26" class='notebook_page'><p>
+
<p>
-
&#9679; Insert 2: Term-Term-pLuxR-LasI-RBS-RFP
+
    LuxR-DT: 15 colonies, picked 1 colony
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>August 13</strong>
+
    pLux-TetR: 35 colonies, picked 1
</p>
</p>
 +
<br/>
<p>
<p>
-
Innoculation
+
    pTet-LasI: Negative control growth; unsuccessful
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LasI
+
    LasR-DT: 25 colonies, picked 1
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LuxI
+
    pLas-LacI: 0 growth
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>August 14</strong>
+
    pLac-LuxI: 15 colonies, picked 1
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
Glycerol Stock and Plasmid isolation
+
    Ligation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LasR-Term-Term-pLasR-RBS-CFP: Concentration: 95.4
+
    LasI into pTet
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP: Concentration: 108.3
+
    Transformation
</p>
</p>
 +
<br/>
<p>
<p>
-
Digestion
+
    pTet-LasI
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LasR-Term-Term-pLasR-RBS-CFP: bad results
+
    pLas-LacI
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP: bad results
+
    Inoculations
</p>
</p>
 +
<br/>
<p>
<p>
-
Parts Recieved
+
    Chlorabackbone
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; BBa_K091159
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; BBa_K091002
+
    August 16
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; BBa_I13453
+
    Colonies
 +
</div>
 +
<div id="page48" class='notebook_page'>
 +
 
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-LuxI:
 +
    </li>
 +
    <li>
 +
        pLac-LasI
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-LasR-Term-Term-pLasR-RBS-CFP: 2 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP: 0 colonies seen
 +
    </li>
 +
    <li>
 +
        LuxS: 200 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        LuxS: 100 colonies seen: 1 colony picked
 +
    </li>
 +
</ul>
 +
<p>
 +
    Glycerol Stock and plasmid isolation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pBad-LsrK: Concentration: 64.2
 +
    </li>
 +
    <li>
 +
        pTetR-LasR-Term-Term-pLasR-RBS-CFP: Concentration:
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP: Concentration:
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-LasR-Term-Term-pLasR-RBS-CFP (E/P)
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP (E/P)
 +
    </li>
 +
    <li>
 +
        pBad-LsrA (S/P) 1749
 +
    </li>
 +
    <li>
 +
        pLsrA-LuxI-RBS-YFP (X/P)
 +
    </li>
 +
</ul>
</div>
</div>
 +
<div id="page49" class='notebook_page'>
 +
<p>
 +
    Testing
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Pop2 (w/ TT) / pTetR-LuxI (6 hours of incubation)
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            100/0
 +
        </li>
 +
        <li>
 +
            100/200
 +
        </li>
 +
        <li>
 +
            100/400
 +
        </li>
 +
        <li>
 +
            100/600
 +
        </li>
 +
        <li>
 +
            Results: After 6 and 24 hours: no considerable difference
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        Pop3 (w/ TT) / pLac-LasI (6 hours of incubation)
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            100/0
 +
        </li>
 +
        <li>
 +
            100/200
 +
        </li>
 +
        <li>
 +
            100/400
 +
        </li>
 +
        <li>
 +
            100/600
 +
        </li>
 +
        <li>
 +
            Results: After 6 and 24 hours: no considerable difference
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        Pop3 (w/ TT) / Pop2 (w/ TT) / pTetR-LuxI (6 hours of incubation)
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            100/0
 +
        </li>
 +
        <li>
 +
            100/200
 +
        </li>
 +
</div>
 +
<div id="page50" class='notebook_page'>
-
<div id="page27" class='notebook_page'><p>
+
        <li>
-
&#9679; BBa_B0034
+
            100/400
 +
        </li>
 +
        <li>
 +
            100/600
 +
        </li>
 +
        <li>
 +
            Results: After 6 and 24 hours: no considerable difference
 +
        </li>
 +
    </ul>
 +
</ul>
 +
<p>
 +
    Digestion
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        population 3 (oscillator)
 +
    </li>
 +
    <li>
 +
        population 2 (oscillator)
 +
    </li>
 +
    <li>
 +
        pBad-&gt;lsrK
 +
    </li>
 +
    <li>
 +
        pLsrA-LuxI-RBS-YFP
 +
    </li>
 +
    <li>
 +
        Term-Term (test)
 +
    </li>
 +
</ul>
<p>
<p>
-
Streaking
+
    Gel Extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pBad-lsrK: Concentration: 53.8
 +
    </li>
 +
    <li>
 +
        pLsrA-LuxI-RBS-YFP: Concentration: 20.0
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; BBa_K091159 pBad-LsrA
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Term-term
 +
    </li>
 +
</ul>
 +
<br/>
 +
 +
</div>
 +
<div id="page51" class='notebook_page'>
<p>
<p>
-
Ligation repeat
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector1: pTetR-LasR
+
    August 16, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert1: Term-Term-pLasR-RBS-CFP
+
    Miniprep/Glycerol Stock
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector2: pLac-LuxR
+
    LuxR-DT
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert 2: Term-Term-pLuxR-LasI-RBS-RFP
+
    pLux-TetR
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>August 15</strong>
+
    LasR-DT
</p>
</p>
 +
<br/>
<p>
<p>
-
Innoculation
+
    pLac-LacI
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LuxI
+
    Chlorabackbone
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LasI
+
    Transformation Results
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LasR-Term-Term-pLasR-RBS-CFP
+
    Negative controls had too much growth; results thrown out
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP
+
    Digestion/Extraction
</p>
</p>
 +
<br/>
<p>
<p>
-
Colonies picked
+
    LuxR-DT: S and P
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pBad-LsrA: 30 colonies seen: 1 picked
+
    pLux-TetR: X and P
</p>
</p>
 +
<br/>
 +
<p>
 +
    Chlorabackbone: E and P
 +
</p>
 +
<br/>
 +
<p>
 +
    Ligations/Transformation
 +
</p>
 +
<br/>
 +
<p>
 +
    LuxR-DT and pLux-TetR
</div>
</div>
 +
<div id="page52" class='notebook_page'>
 +
</p>
 +
<br/>
 +
<p>
 +
    Chlorabackbone with Toggle Switch
 +
</p>
 +
<br/>
 +
<br/>
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
<br/>
 +
<p>
 +
    August 17
 +
</p>
 +
<br/>
 +
<p>
 +
    Glycerol Stock
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Population 3 w/ Term-Term
 +
    </li>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
</ul>
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Population 3 w/ Term-Term: Concentration: 126.7
 +
    </li>
 +
    <li>
 +
        LuxS: Concentration: 40.2
 +
    </li>
 +
    <li>
 +
        LuxS: Concentration: 52.4
 +
    </li>
 +
    <li>
 +
        Term-Term: Concentration: 100.8
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion and Gel Extraction
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Population 3 w/ Term-Term: Concentration: gel not used
 +
    </li>
 +
    <li>
 +
        LuxS: Concentration: gel not used
 +
    </li>
 +
    <li>
 +
        LuxS: Concentration: gel not used
 +
    </li>
 +
</div>
 +
<div id="page53" class='notebook_page'>
-
<div id="page28" class='notebook_page'><p>
+
    <li>
-
Testing
+
        Term-Term: Concentration: gel not used
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector 1: pLac
 +
    </li>
 +
    <li>
 +
        Insert 1: lasI-RBS-RFP
 +
    </li>
 +
    <li>
 +
        Vector 2: pLasR
 +
    </li>
 +
    <li>
 +
        Insert 2: LuxI-RBS-YFP
 +
    </li>
 +
    <li>
 +
        Vector 3: pbad-LsrK
 +
    </li>
 +
    <li>
 +
        Insert 3: pLsrA-&gt;YFP
 +
    </li>
 +
    <li>
 +
        Vector 4: pLac-&gt;LuxR
 +
    </li>
 +
    <li>
 +
        Insert 4: pLuxR-RFP
 +
    </li>
 +
    <li>
 +
        Vector 5: pTetR-LasR
 +
    </li>
 +
    <li>
 +
        Insert 5: pLasR-&gt;CFP
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; Pop2 (w/o TT) / pTetR-LuxI (48 hours of incubation)
+
    Transformation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        BBa_C0080: araC
 +
    </li>
 +
    <li>
 +
        BBa_J24674: pBad-TetR
 +
    </li>
 +
    <li>
 +
        BBa_I14015: pLas-TetO
 +
    </li>
 +
</div>
 +
<div id="page54" class='notebook_page'>
 +
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9675; 100/0 no results
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 100/100 no results
+
    August 17, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 100/200 slight RFP detected no increase of RFP from 100/0
+
    Ligation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Pop3 (w/o TT) / pLac-LasI (48 hours of incubation)
+
    pLas with LacI
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 100/0 no results
+
    pTet with LasI
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9675; 100/100 no results
+
    Inoculations
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 100/200 slight RFP detected no increase of CFP from 100/0
+
    pLas
</p>
</p>
 +
<br/>
<p>
<p>
-
Streaking
+
    LacI
</p>
</p>
 +
<br/>
<p>
<p>
-
BBa_K091109
+
    pTet
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; SP3011&gt;1D LuxS
+
    pLac
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; SP4011&gt;1D LuxS
+
    GFP
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>August 16</strong>
+
    RFP-LVA
</p>
</p>
 +
<br/>
<p>
<p>
-
Colonies
+
    LasI
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LuxI:
+
    Transformation
</p>
</p>
 +
<br/>
 +
<p>
 +
    Bba_K081014 Kit Plate 3 Well 12O Resistance AK; RFP
 +
</p>
 +
<br/>
 +
<br/>
</div>
</div>
 +
<div id="page55" class='notebook_page'>
-
 
+
<p>
-
<div id="page29" class='notebook_page'><p>
+
    Triple Population and AND Gate Projects
-
&#9679; pLac-LasI
+
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LasR-Term-Term-pLasR-RBS-CFP: 2 colonies seen: 1 colony picked
+
    August 18
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP: 0 colonies seen
+
    Innoculations
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
    <li>
 +
        pLac-lasI
 +
    </li>
 +
    <li>
 +
        pop 2 w/TT (oscillator)
 +
    </li>
 +
    <li>
 +
        pop 3 w/TT (oscillator)
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: 200 colonies seen: 1 colony picked
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: 100 colonies seen: 1 colony picked
+
    August 18, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
Glycerol Stock and plasmid isolation
+
    Colony Picking
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pBad-LsrK: Concentration: 64.2
+
    LuxR-DT-pLux-TetR: 30 colonies, 1 picked
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LasR-Term-Term-pLasR-RBS-CFP: Concentration:
+
    Toggle Switch in Chlorabackbone: 4 colonies, 1 picked
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP: Concentration:
+
    pLas with LacI: negative control growth; results thrown out
</p>
</p>
 +
<br/>
<p>
<p>
-
Digestion
+
    pTet with LasI: negative control growth; results thrown out
</p>
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    RFP: 20 colonies, 1 picked
 +
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LasR-Term-Term-pLasR-RBS-CFP (E/P)
+
    Glycerol Stock
</p>
</p>
 +
<br/>
 +
</div>
 +
<div id="page56" class='notebook_page'>
 +
<p>
<p>
-
&#9679; pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP (E/P)
+
    LuxR-DT-pLux-TetR
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pBad-LsrA (S/P) 1749
+
    Minipreps
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLsrA-LuxI-RBS-YFP (X/P)
+
    LuxR-DT-pLux-TetR
</p>
</p>
 +
<br/>
 +
<p>
 +
    pLas
 +
</p>
 +
<br/>
 +
<p>
 +
    LacI
 +
</p>
 +
<br/>
 +
<p>
 +
    pTet
 +
</p>
 +
<br/>
 +
<p>
 +
    pLac
 +
</p>
 +
<br/>
 +
<p>
 +
    GFP
 +
</p>
 +
<br/>
 +
<p>
 +
    LasI
 +
</p>
 +
<br/>
 +
<br/>
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
<br/>
</div>
</div>
 +
<div id="page57" class='notebook_page'>
 +
<p>
 +
    August 19
 +
</p>
 +
<br/>
 +
<p>
 +
    Glycerol Stocks
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-lasI-RBS-RFP
 +
    </li>
 +
    <li>
 +
        pLasR-LuxI-RBS-YFP
 +
    </li>
 +
    <li>
 +
        pBad-LsrK-plsrA-LuxI-RBS-YFP
 +
    </li>
 +
    <li>
 +
        plac-LuxR-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-pLasR-RBS-CFP
 +
    </li>
 +
    <li>
 +
        araC
 +
    </li>
 +
    <li>
 +
        pBad-TetR
 +
    </li>
 +
    <li>
 +
        pLas-TetO
 +
    </li>
 +
</ul>
 +
<p>
 +
    Oscillator Population Tests (population with induction plasmid)
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pop2 0:1 - some fluorescence
 +
    </li>
 +
    <li>
 +
        pop2 75:1 - no visual difference
 +
    </li>
 +
    <li>
 +
        pop3 0:1 - some fluorescence
 +
    </li>
 +
    <li>
 +
        pop3 75:1 - no visual difference
 +
    </li>
 +
    <li>
 +
        pop2 and pop3 0:1 - little fluorescence
 +
    </li>
 +
</div>
 +
<div id="page58" class='notebook_page'>
-
<div id="page30" class='notebook_page'><p>
+
    <li>
-
Testing
+
        pop2 and pop3 75:1 - a visual increase of both RFP and CFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-lasI-RBS-RFP: Concentration: 115.8
 +
    </li>
 +
    <li>
 +
        pLasR-LuxI-RBS-YFP: Concentration: 93.1
 +
    </li>
 +
    <li>
 +
        pBad-LsrK-plsrA-LuxI-RBS-YFP: Concentration: 128.6
 +
    </li>
 +
    <li>
 +
        plac-LuxR-pLuxR-LasI-RBS-RFP: Concentration: 105.9
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-pLasR-RBS-CFP: Concentration: 122.3
 +
    </li>
 +
    <li>
 +
        araC: Concentration: 114.0
 +
    </li>
 +
    <li>
 +
        pBad-TetR: Concentration: 315.1
 +
    </li>
 +
    <li>
 +
        pLas-TetO: Concentration: 102.3
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; Pop2 (w/ TT) / pTetR-LuxI (6 hours of incubation)
+
    Digestion and gel extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        ad-LsrK-plsrA-LuxI-RBS-YFP: Concentration:
 +
    </li>
 +
    <li>
 +
        plac-LuxR-pLuxR-LasI-RBS-RFP: Concentration:
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-pLasR-RBS-CFP: Concentration:
 +
    </li>
 +
    <li>
 +
        araC: Concentration:
 +
    </li>
 +
    <li>
 +
        pBad-TetR: Concentration:
 +
    </li>
 +
</div>
 +
<div id="page59" class='notebook_page'>
 +
 +
    <li>
 +
        pLas-TetO: Concentration:
 +
    </li>
 +
    <li>
 +
        LasR-TT: Concentration: 10.0
 +
    </li>
 +
    <li>
 +
        LuxS: Concentration: 3.1
 +
    </li>
 +
    <li>
 +
        LuxS: Concentration: 5.5
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9675; 100/0
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector 1: pyear
 +
    </li>
 +
    <li>
 +
        Insert 1: LuxS
 +
    </li>
 +
    <li>
 +
        Vector 2: pyear
 +
    </li>
 +
    <li>
 +
        Insert 2: LuxS
 +
    </li>
 +
    <li>
 +
        Vector 3: pTetR
 +
    </li>
 +
    <li>
 +
        Insert 3:lasR-TT
 +
    </li>
 +
    <li>
 +
        Vector 4: pLac-LuxR
 +
    </li>
 +
    <li>
 +
        Insert 4: TT
 +
    </li>
 +
    <li>
 +
        Vector 5: pLuxR
 +
    </li>
 +
    <li>
 +
        Insert 5: arac
 +
    </li>
 +
    <li>
 +
        Vector 6: pBad-TetR
 +
    </li>
 +
</div>
 +
<div id="page60" class='notebook_page'>
 +
 +
    <li>
 +
        Insert 6: TT
 +
    </li>
 +
    <li>
 +
        Vector 7: plasR-TetO
 +
    </li>
 +
    <li>
 +
        Insert 7: RBS-CFP
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9675; 100/200
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 100/400
+
    August 19, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 100/600
+
    Glycerol Stock/Plasmid Isolation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; Results: After 6 and 24 hours: no considerable difference
+
    RFP
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Pop3 (w/ TT) / pLac-LasI (6 hours of incubation)
+
    Toggle Switch in Chlorabackbone
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9675; 100/0
+
    Inoculation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 100/200
+
    RFP-LVA
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9675; 100/400
+
    Digestion/Extraction
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 100/600
+
    pLac: S and P
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; Results: After 6 and 24 hours: no considerable difference
+
    LuxR-DT-pLux-TetR: X and P
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Pop3 (w/ TT) / Pop2 (w/ TT) / pTetR-LuxI (6 hours of incubation)
+
    pTet: S and P
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 100/0
+
    LasI: X and P
</p>
</p>
-
 
+
<br/>
<p>
<p>
-
&#9675; 100/200
+
    LasR-DT: X and P
</p>
</p>
 +
<br/>
</div>
</div>
 +
<div id="page61" class='notebook_page'>
-
<div id="page31" class='notebook_page'><p>
+
<p>
-
&#9675; 100/400
+
    LacI: S and P
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 100/600
+
    RFP: X and P
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9675; Results: After 6 and 24 hours: no considerable difference
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Digestion
+
    August 20
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; population 3 (oscillator)
+
    Colonies picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pyear-LuxR: 1 picked
 +
    </li>
 +
    <li>
 +
        pyear-LuxR: 1 picked
 +
    </li>
 +
    <li>
 +
        pTet-lasR-TT: 1 picked
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-TT: 1 picked
 +
    </li>
 +
    <li>
 +
        pBad-TetR-TT: 1 picked
 +
    </li>
 +
    <li>
 +
        pLuxR-arac: 1 picked
 +
    </li>
 +
    <li>
 +
        pLasR-TetO-RBS-CFP: 1 picked after being repeated
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; population 2 (oscillator)
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
    <li>
 +
        pLac-lasI
 +
    </li>
 +
    <li>
 +
        pop3 w/TT
 +
    </li>
 +
 +
</div>
 +
<div id="page62" class='notebook_page'>
 +
 +
    <li>
 +
        pop2 w/TT
 +
    </li>
 +
    <li>
 +
        pop3 w/o TT
 +
    </li>
 +
    <li>
 +
        pop2 w/o TT
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; pBad-&gt;lsrK
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLsrA-LuxI-RBS-YFP
+
    August 20, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Term-Term (test)
+
    Ligations/Transformations
</p>
</p>
 +
<br/>
<p>
<p>
-
Gel Extraction
+
    LuxR-DT-pLux-TetR into pLac
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pBad-lsrK: Concentration: 53.8
+
    LasI into pTet
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLsrA-LuxI-RBS-YFP: Concentration: 20.0
+
    LasR-DT into pTet
</p>
</p>
 +
<br/>
<p>
<p>
-
Innoculation
+
    RFP into LacI
</p>
</p>
 +
<br/>
 +
<br/>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; Term-term
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>August 17</strong>
+
    August 21
</p>
</p>
 +
<br/>
<p>
<p>
-
Glycerol Stock
+
    Glycerol Stock and plasmid isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pyear-LuxR: Concentration: 40.0
 +
    </li>
 +
    <li>
 +
        pyear-LuxR: Concentration: 75.0
 +
    </li>
 +
    <li>
 +
        pTet-lasR-T: Concentration: 127.6
 +
    </li>
</div>
</div>
 +
<div id="page63" class='notebook_page'>
-
<div id="page32" class='notebook_page'><p>
+
    <li>
-
&#9679; Population 3 w/ Term-Term
+
        pLac-LuxR-TT: Concentration: 123.5
 +
    </li>
 +
    <li>
 +
        pBad-TetR-TT: Concentration: 141.0
 +
    </li>
 +
    <li>
 +
        pLuxR-arac: Concentration: 84.0
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion and gel extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLasR-LuxI-YFP: gel not used
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-TT: gel not used
 +
    </li>
 +
    <li>
 +
        pLuxR-araC: gel not used
 +
    </li>
 +
    <li>
 +
        pyear-LuxS: gel not used
 +
    </li>
 +
    <li>
 +
        pyear-LuxS: gel not used
 +
    </li>
 +
    <li>
 +
        pop1 (test)
 +
    </li>
 +
    <li>
 +
        pop2 (test)
 +
    </li>
 +
    <li>
 +
        pop3 (test)
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; LuxS
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pyear-LuxS
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; LuxS
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector 1: pTet-LasR-TT
 +
    </li>
 +
</div>
 +
<div id="page64" class='notebook_page'>
 +
 +
    <li>
 +
        Insert 1: pLasR-LuxI-RBS-YFP
 +
    </li>
 +
    <li>
 +
        Vector 2: pLuxR-araC
 +
    </li>
 +
    <li>
 +
        Insert 2: lasR-TT
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
Plasmid Isolation
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Population 3 w/ Term-Term: Concentration: 126.7
+
    August 21, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: Concentration: 40.2
+
    Colony Pickings
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: Concentration: 52.4
+
    pLac-LuxR-DT-pLux-TetR: 24 colonies, 1 picked
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Term-Term: Concentration: 100.8
+
    pTet-LasI: 30 colonies, 3 picked
</p>
</p>
 +
<br/>
<p>
<p>
-
Digestion and Gel Extraction
+
    pTet-LasR-DT: 100 colonies, 2 picked
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Population 3 w/ Term-Term: Concentration: gel not used
+
    LacI-RFP: 24 colonies, 2 picked
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: Concentration: gel not used
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LuxS: Concentration: gel not used
+
    August 22
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Term-Term: Concentration: gel not used
+
    Digestion (test)
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        population 2 (oscillator)
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            undigested
 +
        </li>
 +
        <li>
 +
            EcoRI
 +
        </li>
 +
</div>
 +
<div id="page65" class='notebook_page'>
 +
 +
        <li>
 +
            XbaI
 +
        </li>
 +
        <li>
 +
            Spe1
 +
        </li>
 +
        <li>
 +
            Pst1
 +
        </li>
 +
        <li>
 +
            E/P
 +
        </li>
 +
        <li>
 +
            X/S
 +
        </li>
 +
    </ul>
 +
</ul>
<p>
<p>
-
Ligation
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        population 1 (oscillator)
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; Vector 1: pLac
+
    Colonies picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-lasR-TT-pLasR-LuxI-RBS-YFP
 +
    </li>
 +
    <li>
 +
        pLuxR-araC-LasR-TT
 +
    </li>
 +
    <li>
 +
        pLasR-TetO-RBS-CFP (2 colonies picked)
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; Insert 1: lasI-RBS-RFP
+
    Transformation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR
 +
    </li>
 +
</ul>
 +
<p>
 +
    Innoculation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
</div>
</div>
 +
<div id="page66" class='notebook_page'>
-
<div id="page33" class='notebook_page'><p>
+
<p>
-
&#9679; Vector 2: pLasR
+
    Population Testing Prep (pop2, pop3, pop2+pop3)
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        2 ml of medium added to each test tube
 +
    </li>
 +
    <li>
 +
        100 microliters of each population added as necessary
 +
    </li>
 +
    <li>
 +
        .75mL of inducing population added as necessary
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            pop2 w/ w/o inducer
 +
        </li>
 +
        <li>
 +
            pop3 w/ w/o inducer
 +
        </li>
 +
        <li>
 +
            pop2+pop3 w/ w/o inducer
 +
        </li>
 +
    </ul>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; Insert 2: LuxI-RBS-YFP
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector 3: pbad-LsrK
+
    August 22, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert 3: pLsrA-&gt;YFP
+
    Glycerol Stock/Miniprep
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector 4: pLac-&gt;LuxR
+
    pLac-LuxR-DT-pLux-TetR
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert 4: pLuxR-RFP
+
    pTet-LasI
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector 5: pTetR-LasR
+
    pTet-LasR-DT
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert 5: pLasR-&gt;CFP
+
    LacI-RFP
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
Transformation
+
    Digestion/Extraction:
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; BBa_C0080: araC
+
    pLac-LuxR-DT-pLux-TetR: S and P
</p>
</p>
 +
<br/>
 +
</div>
 +
<div id="page67" class='notebook_page'>
 +
<p>
<p>
-
&#9679; BBa_J24674: pBad-TetR
+
    GFP: X and P
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; BBa_I14015: pLas-TetO
+
    LacI-RFP: S and P
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>August 18</strong>
+
    pLac-LuxI: X and P
</p>
</p>
 +
<br/>
<p>
<p>
-
Innoculations
+
    Test Digestions
 +
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    pTet-LasI: E and P
</p>
</p>
<p>
<p>
-
&#9679; pTetR-LuxI
+
    pTet-LasR-DT: E and P
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLac-lasI
+
    Ligations/Transformations
</p>
</p>
-
</div>
+
<br/>
-
 
+
<p>
-
<div id="page34" class='notebook_page'><p>
+
    GFP into pLac-LuxR-DT-pLux-TetR
-
&#9679; pop 2 w/TT (oscillator)
+
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pop 3 w/TT (oscillator)
+
    pLac-LuxI into LacI-RFP
</p>
</p>
 +
<br/>
 +
<br/>
 +
<p>
 +
    Inoculations
 +
</p>
 +
<br/>
 +
<p>
 +
    RFP-LVA
 +
</p>
 +
<br/>
 +
<p>
 +
    pTet-GFP
 +
</p>
 +
<br/>
 +
<p>
 +
    RBS-YFP
 +
</p>
 +
<br/>
 +
<br/>
 +
<p>
 +
    August 22
 +
</p>
 +
<br/>
 +
<p>
 +
    Digestion (test)
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        population 2 (oscillator)
 +
    </li>
</div>
</div>
 +
<div id="page68" class='notebook_page'>
-
<div id="page35" class='notebook_page'><p>
+
    <ul>
-
<strong>August 19</strong>
+
        <li>
 +
            undigested
 +
        </li>
 +
        <li>
 +
            EcoRI
 +
        </li>
 +
        <li>
 +
            XbaI
 +
        </li>
 +
        <li>
 +
            Spe1
 +
        </li>
 +
        <li>
 +
            Pst1
 +
        </li>
 +
        <li>
 +
            E/P
 +
        </li>
 +
        <li>
 +
            X/S
 +
        </li>
 +
    </ul>
 +
</ul>
 +
<p>
 +
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        population 1 (oscillator)
 +
    </li>
 +
</ul>
<p>
<p>
-
Glycerol Stocks
+
    Colonies picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-lasR-TT-pLasR-LuxI-RBS-YFP
 +
    </li>
 +
    <li>
 +
        pLuxR-araC-LasR-TT
 +
    </li>
 +
    <li>
 +
        pLasR-TetO-RBS-CFP (2 colonies picked)
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pLac-lasI-RBS-RFP
+
    Transformation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pLasR-LuxI-RBS-YFP
+
    Innoculation
</p>
</p>
 +
<br/>
 +
</div>
 +
<div id="page69" class='notebook_page'>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pBad-LsrK-plsrA-LuxI-RBS-YFP
+
    Population Testing Prep (pop2, pop3, pop2+pop3)
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        2 ml of medium added to each test tube
 +
    </li>
 +
    <li>
 +
        100 microliters of each population added as necessary
 +
    </li>
 +
    <li>
 +
        .75mL of inducing population added as necessary
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            pop2 w/ w/o inducer
 +
        </li>
 +
        <li>
 +
            pop3 w/ w/o inducer
 +
        </li>
 +
        <li>
 +
            pop2+pop3 w/ w/o inducer
 +
        </li>
 +
    </ul>
 +
</ul>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; plac-LuxR-pLuxR-LasI-RBS-RFP
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-lasR-pLasR-RBS-CFP
+
    August 23, 2012
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; araC
+
    Colony Picking
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pBad-TetR
+
    LacI-RFP-ST-pLac-LuxI; 40 colonies, 1 picked
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLas-TetO
+
    LacI-RFP-ST-pLac-LuxI; 30 colonies, 1 picked
</p>
</p>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
Oscillator Population Tests (population with induction plasmid)
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pop2 0:1 - some fluorescence
+
    August 23
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pop2 75:1 - no visual difference
+
    Glycerol Stock
</p>
</p>
 +
<br/>
 +
 +
</div>
 +
<div id="page70" class='notebook_page'>
 +
<ul>
 +
    <li>
 +
        pLasR-TetO-RBS-CFP (X2)
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-TT-pLasR-LuxI-RBS-YFP
 +
    </li>
 +
    <li>
 +
        pLuxR-araC-LasR-TT
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pop3 0:1 - some fluorescence
+
    Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLasR-TetO-RBS-CFP: Concentration: 113.7
 +
    </li>
 +
    <li>
 +
        pLasR-TetO-RBS-CFP: Concentration: 112.0
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-lasR-TT-pLasR-LuxI-RBS-YFP: Concentration: 167.0
 +
    </li>
 +
    <li>
 +
        pLuxR-araC-LasR-TT: Concentration: 104.2
 +
    </li>
 +
    <li>
 +
        pLac-lasI-RBS-RFP: Concentration: 37.0
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pop3 75:1 - no visual difference
+
    Colonies picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pop2 and pop3 0:1 - little fluorescence
+
    Population Testing from 8/22 prep after 24 hours
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        500 microliters from test tube of populations put in microcentrifuge tubes
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <ul>
 +
        <li>
 +
            pop2 w/: slight increase of RFP
 +
        </li>
 +
        <li>
 +
            pop2 w/o: some RFP
 +
        </li>
 +
        <li>
 +
            pop3 w/: no significant increase of CFP
 +
        </li>
 +
</div>
</div>
-
<div id="page36" class='notebook_page'><p>
+
<div id="page71" class='notebook_page'>
-
&#9679; pop2 and pop3 75:1 - a visual increase of both RFP and CFP
+
        <li>
-
</p>
+
            pop3 w/o: some CFP
 +
        </li>
 +
        <li>
 +
            pop2+pop3 w/: no significant increase of RFP/CFP
 +
        </li>
 +
        <li>
 +
            pop2+pop3 w/o: some RFP/CFP
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        Results:
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            test tubes were checked after 24 hours
 +
        </li>
 +
        <ul>
 +
            <li>
 +
                pop2 showed a slight increase in RFP production when it was induced in comparison to its uninduced control
 +
            </li>
 +
            <li>
 +
                both pop3 and pop2+pop3 showed no significant differences between its positive and negative controls
 +
            </li>
 +
        </ul>
 +
        <li>
 +
            after 48 hours
 +
        </li>
 +
        <ul>
 +
            <li>
 +
                pop2 showed increase in RFP production
 +
            </li>
 +
            <li>
 +
                pop3 showed no significant increase in CFP production
 +
            </li>
 +
            <li>
 +
                pop2+pop3 showed a sig increase in RFP production but no CFP when it was induced compared to its negative control
 +
            </li>
 +
        </ul>
 +
    </ul>
 +
</ul>
<p>
<p>
-
Plasmid Isolation
+
    Digestion
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        plac-LuxR(S/P): Concentration: 19.7
 +
    </li>
 +
</div>
 +
<div id="page72" class='notebook_page'>
 +
    <li>
 +
        pTetR-LasR (E/P): Concentration: gel not used
 +
    </li>
 +
    <li>
 +
        pLasR-RBS-CFP (X/P): Concentration: gel not used
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-TT (S/P): Concentration: 24.0
 +
    </li>
 +
    <li>
 +
        pBad-TetR-TT (S/P): Concentration: 15.4
 +
    </li>
 +
    <li>
 +
        pTetR-LasR-TT-pLasR-LuxI-RBS-YFP (E/P): Concentration: 12.2
 +
    </li>
 +
    <li>
 +
        pLuxR-araC-lasR-TT (X/P): Concentration: gel not used
 +
    </li>
 +
    <li>
 +
        pLasR-TetO-RBS-CFP (X/P): Concentration: 5.2
 +
    </li>
 +
    <li>
 +
        pLasR-TetO-RBS-CFP (X/P): Concentration: 5.2
 +
    </li>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP (X/P): Concentration: gel not used
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pLac-lasI-RBS-RFP: Concentration: 115.8
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pyear-luxS
 +
    </li>
 +
    <li>
 +
        pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
 +
    </li>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
    <li>
 +
        pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
 +
    </li>
 +
    <li>
 +
        pLac-lasI
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
 +
    </li>
 +
</div>
 +
<div id="page73" class='notebook_page'>
 +
 +
    <li>
 +
        pLuxR-lasI-RBS-RFP
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pLasR-LuxI-RBS-YFP: Concentration: 93.1
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector: pBad-TetR-TT
 +
    </li>
 +
    <li>
 +
        Insert: plasR-TetO-RBS-CFP
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; pBad-LsrK-plsrA-LuxI-RBS-YFP: Concentration: 128.6
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; plac-LuxR-pLuxR-LasI-RBS-RFP: Concentration: 105.9
+
    August 24, 2012
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-lasR-pLasR-RBS-CFP: Concentration: 122.3
+
    Plasmid Isolation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; araC: Concentration: 114.0
+
    LacI-RFP-ST-pLac-LuxI
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pBad-TetR: Concentration: 315.1
+
    Digestion/Extraction
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLas-TetO: Concentration: 102.3
+
    pTet-LasR-DT
</p>
</p>
 +
<br/>
<p>
<p>
-
Digestion and gel extraction
+
    pTet-LasI
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; ad-LsrK-plsrA-LuxI-RBS-YFP: gel not used
+
    LacI-RFP-ST-pLac-LuxI
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; plac-LuxR-pLuxR-LasI-RBS-RFP: gel not used
+
    pLas
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-lasR-pLasR-RBS-CFP: gel not used
+
    Ligation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; araC: gel not used
+
    pTet-LasI into pLac-LuxR-DT-pLux-TetR-GFP-ST
</p>
</p>
 +
<br/>
 +
<br/>
 +
</div>
 +
<div id="page74" class='notebook_page'>
 +
<p>
<p>
<p>
-
&#9679; pBad-TetR: gel not used
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
</div>
</div>
-
 
+
    August 24
-
 
+
-
<div id="page37" class='notebook_page'><p>
+
-
&#9679; pLas-TetO: gel not used
+
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; LasR-TT: Concentration: 10.0
+
    Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pluxR-LasI-RBS-RFP: Concentration: 42.6
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; LuxS: Concentration: 3.1
+
    Digestion
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP (X/P)
 +
    </li>
 +
    <li>
 +
        pLuxR-araC-lasR-TT (X/P)
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; LuxS: Concentration: 5.5
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP
 +
    </li>
 +
    <li>
 +
        pLuxR-araC-LasR-TT
 +
    </li>
 +
    <li>
 +
        pLsrA-LuxI-YFP
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
Ligation
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector 1: pyear
+
    August 25, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert 1: LuxS
+
    Glycerol Stock/Plasmid Isolation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector 2: pyear
+
    LacI-RFP-ST-pLac-LuxI
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; Insert 2: LuxS
+
    Digestion/Extraction
</p>
</p>
 +
<br/>
 +
</div>
 +
<div id="page75" class='notebook_page'>
 +
<p>
<p>
<p>
-
&#9679; Vector 3: pTetR
+
    LacI-RFP-ST-pLac-LuxI
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; Insert 3:lasR-TT
+
    Ligation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector 4: pLac-LuxR
+
    LacI-RFP-ST-pLac-LuxI into pLas
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; Insert 4: TT
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector 5: pLuxR
+
    August 25
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert 5: arac
+
    Glycerol Stock
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        plac-LuxR
 +
    </li>
 +
    <li>
 +
        pBad-TetR-TT-plasR-TetO-RBS-CFP
 +
    </li>
 +
    <li>
 +
        pBad-TetR-TT-plasR-TetO-RBS-CFP
 +
    </li>
 +
    <li>
 +
        pLuxR-araC-lasR-TT
 +
    </li>
 +
</ul>
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP: Concentration: 89.1
 +
    </li>
 +
    <li>
 +
        pLuxR-araC-LasR-TT: Concentration: 152.9
 +
    </li>
 +
    <li>
 +
        pBad-TetR-TT-plasR-TetO-RBS-CFP: Concentration: 200.0
 +
    </li>
 +
    <li>
 +
        pBad-TetR-TT-plasR-TetO-RBS-CFP: Concentration: 156.0
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; Vector 6: pBad-TetR
+
    Digestion and Gel Extraction
</p>
</p>
 +
<br/>
</div>
</div>
 +
<div id="page76" class='notebook_page'>
 +
<p>
-
<div id="page38" class='notebook_page'><p>
+
<ul>
-
&#9679; Insert 6: TT
+
    <li>
-
</p>
+
        pLuxR-LasI-RBS-RFP: (X/P): Concentration: 9.5
 +
    </li>
 +
    <li>
 +
        pLuxR-araC-LasR-TT: (X/P): gel not used
 +
    </li>
 +
    <li>
 +
        pBad-TetR-TT-plasR-TetO-RBS-CFP: (X/P): gel not used
 +
    </li>
 +
    <li>
 +
        pBad-TetR-TT-plasR-TetO-RBS-CFP: (X/P): gel not used
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; Vector 7: plasR-TetO
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector1: pLac-LuxR-TT
 +
    </li>
 +
    <li>
 +
        Insert1: pLuxR-LasI-RBS-RFP
 +
    </li>
 +
    <li>
 +
        Vector2: pTetR-LasR
 +
    </li>
 +
    <li>
 +
        Insert2: pLasR-CFP
 +
    </li>
 +
    <li>
 +
        Vector3: pLac-LuxR-TT
 +
    </li>
 +
    <li>
 +
        Insert3: pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; Insert 7: RBS-CFP
+
    Primer Dilution
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        I3.PsB1A2.R +661ul NFH2O -&gt; 1:10 Dulition
 +
    </li>
 +
    <li>
 +
        I3.DT.F +634ul NFH2O -&gt; 1:10 Dulition
 +
    </li>
 +
    <li>
 +
        I3.FT.M +700ul NFH2O -&gt; 1:10 Dulition
 +
    </li>
 +
</ul>
<p>
<p>
-
<strong>August 20</strong>
+
    Colony PCR
</p>
</p>
 +
<br/>
 +
</div>
 +
<div id="page77" class='notebook_page'>
<p>
<p>
-
Colonies picked
+
<ul>
-
</p>
+
    <li>
 +
        pBad-TetR-TT-plasR-TetO-RBS-CFP
 +
    </li>
 +
    <li>
 +
        pLuxR-araC-LasR-TT
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            Primer 1: I3.24.SP1
 +
        </li>
 +
        <li>
 +
            Primer 2: I3.PSB1A2.R
 +
        </li>
 +
    </ul>
 +
</ul>
<p>
<p>
-
&#9679; pyear-LuxR: 1 picked
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pyear-luxS
 +
    </li>
 +
    <li>
 +
        pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
 +
    </li>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
    <li>
 +
        pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
 +
    </li>
 +
    <li>
 +
        pLac-lasI
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
 +
    </li>
 +
    <li>
 +
        pLuxR-lasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
</div>
 +
<div id="page78" class='notebook_page'>
 +
<p>
 +
Triple Population and AND Gate Projects
<p>
<p>
-
&#9679; pyear-LuxR: 1 picked
+
    August 26, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTet-lasR-TT: 1 picked
+
    Colony Picking
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-TT: 1 picked
+
    pLas-LacI-RFP-ST-pLac-LuxI; 50 colonies, 1 picked
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pBad-TetR-TT: 1 picked
+
    Inoculation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-arac: 1 picked
+
    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLasR-TetO-RBS-CFP: 1 picked after being repeated
+
    pLas-LacI-RFP-ST-pLac-LuxI
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
Innoculation
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LuxI
+
    August 26
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-lasI
+
    Colonies Picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLuxR-araC-LasR-TT: 1 colony picked
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pop3 w/TT
+
    Population testing
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        2ml broth, 1ml induction, 100ul population, 30 hour incubation
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            pop1+2 (+/-): no fluorescence seen
 +
        </li>
 +
        <li>
 +
            pop2+3 (+/-): no fluorescence seen
 +
        </li>
 +
        <li>
 +
            pop1+2+3 (+/-): no fluorescence seen
 +
        </li>
</div>
</div>
 +
<div id="page79" class='notebook_page'>
-
<div id="page39" class='notebook_page'><p>
+
    </ul>
-
&#9679; pop2 w/TT
+
</ul>
 +
<br/>
 +
<p>
 +
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pop3 w/o TT
+
    August 27, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pop2 w/o TT
+
    Glycerol Stock/Plasmid Isolation
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>August 21</strong>
+
    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP
</p>
</p>
 +
<br/>
<p>
<p>
-
Glycerol Stock and plasmid isolation
+
    pLas-LacI-RFP-ST-pLac-LuxI
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pyear-LuxR: Concentration: 40.0
+
    PCR
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pyear-LuxR: Concentration: 75.0
+
    Add second LacO site to pLac
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pTet-lasR-T: Concentration: 127.6
+
    Digestion/Extraction
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-TT: Concentration: 123.5
+
    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pBad-TetR-TT: Concentration: 141.0
+
    pLas-LacI-RFP-ST-pLac-LuxI
</p>
</p>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-arac: Concentration: 84.0
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Digestion and gel extraction
+
    August 27
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLasR-LuxI-YFP: gel not used
+
    Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLuxR-araC-LaR-TT: Concentration: 164.3
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pTetR-lasR-TT: gel not used
+
    Digestion and Gel extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLuxR-araC-LaR-TT (X/P): Concentration: 55.1
 +
    </li>
 +
</ul>
 +
</div>
 +
<div id="page80" class='notebook_page'>
 +
<p>
<p>
-
&#9679; pLuxR-araC: gel not used
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pyear-luxS
 +
    </li>
 +
    <li>
 +
        pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
 +
    </li>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
    <li>
 +
        pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
 +
    </li>
 +
    <li>
 +
        pLac-lasI
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
 +
    </li>
 +
    <li>
 +
        pLuxR-lasI-RBS-RFP
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pyear-LuxS: gel not used
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector1: pBad-tetR-TT
 +
    </li>
 +
    <li>
 +
        Insert1: pLasR-TetO-RBS-CFP
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            0:1 20 colonies seen
 +
        </li>
 +
        <li>
 +
            2:1 50 colonies seen
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        Vector2:pLac-LuxR-TT
 +
    </li>
 +
    <li>
 +
        Insert2: pLuXR-araC-lasR-TT
 +
    </li>
</div>
</div>
-
 
+
<div id="page81" class='notebook_page'>
-
<div id="page40" class='notebook_page'><p>
+
    <ul>
-
&#9679; pyear-LuxS: gel not used
+
        <li>
 +
            0:1 5 colonies seen
 +
        </li>
 +
        <li>
 +
            2:1 25 colonies seen
 +
        </li>
 +
    </ul>
 +
</ul>
 +
<br/>
 +
<p>
 +
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pop1 (test)
+
    August 28, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pop2 (test)
+
    Primer Dilutions
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pop3 (test)
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Innoculation
+
    August 28
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pyear-LuxS
+
    Colony PCR
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pBad-tetR-TT-pLasR-TetO-RBS-CFP
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-TT- pLuXR-araC-lasR-TT
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Primer 1: I3.24.SP1
 +
    </li>
 +
    <li>
 +
        Primer 2: I3.PSB1A2.R
 +
    </li>
 +
</ul>
<p>
<p>
-
Ligation
+
    Population Testing
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        200ul medium, 200ul induction, 4ul population, 24 hour incubation
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pop1+2 (-): no fluorescence seen
 +
    </li>
 +
    <li>
 +
        pop1+2 (+): some sign of RFP/YFP
 +
    </li>
 +
</div>
 +
<div id="page82" class='notebook_page'>
 +
    <li>
 +
        pop2+3 (-): no fluorescence seen
 +
    </li>
 +
    <li>
 +
        pop2+3 (+): no fluorescence seen
 +
    </li>
 +
    <li>
 +
        pop1+2+3 (-): no fluorescence seen
 +
    </li>
 +
    <li>
 +
        pop1+2+3 (+): no fluorescence seen
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; Vector 1: pTet-LasR-TT
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector:pLac-LuxR-TT
 +
    </li>
 +
    <li>
 +
        Insert: pLuXR-araC-lasR-TT
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            0:1 0 colonies seen
 +
        </li>
 +
        <li>
 +
            2:1 30 colonies seen
 +
        </li>
 +
        <li>
 +
            4:1 15 colonies seen
 +
        </li>
 +
    </ul>
 +
</ul>
<p>
<p>
-
&#9679; Insert 1: pLasR-LuxI-RBS-YFP
+
    Digestion and Gel Extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pBad-TetR-TT (S/P): Concentration: 88.3
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; Vector 2: pLuxR-araC
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pyear-luxS
 +
    </li>
 +
    <li>
 +
        pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
 +
    </li>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
    <li>
 +
        pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
 +
    </li>
 +
</div>
 +
<div id="page83" class='notebook_page'>
 +
    <li>
 +
        pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; Insert 2: lasR-TT
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>August 22</strong>
+
    August 29, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
Digestion (test)
+
    Ligation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; population 2 (oscillator)
+
    pLas-LacI-RFP-ST-pLac-LuxI into pTet-LasR-DT
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; undigested
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; EcoRI
+
    August 29
</p>
</p>
 +
<br/>
 +
<p>
 +
    Population Test Prep
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        2ml medium, 400ul populations, 1500ul induction
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            pop1+2 (+/-)
 +
        </li>
 +
        <li>
 +
            pop2+3 (+/-)
 +
        </li>
 +
        <li>
 +
            pop1+2+3 (+/-)
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        1ml medium, 200ul populations, 750ul induction
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            pop2+3 (+/-)
 +
        </li>
 +
    </ul>
 +
</ul>
</div>
</div>
-
 
+
<div id="page84" class='notebook_page'>
-
 
+
<p>
-
<div id="page41" class='notebook_page'><p>
+
    Triple Population and AND Gate Projects
-
&#9675; XbaI
+
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; Spe1
+
    August 30
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; Pst1
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector: pBad-TetR-TT
 +
    </li>
 +
    <li>
 +
        Insert: plasR-TetO-CFP
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            0:1 100 Colonies seen
 +
        </li>
 +
        <li>
 +
            1:1 200 Colonies seen
 +
        </li>
 +
        <li>
 +
            4:1 200 Colonies seen
 +
        </li>
 +
    </ul>
 +
</ul>
<p>
<p>
-
&#9675; E/P
+
    Colony PCR
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-TT-pLuXR-araC-lasR-TT (X2)
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Primer 1: I3.24.SP1
 +
    </li>
 +
    <li>
 +
        Primer 2: I3.PSB1A2.R
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9675; X/S
+
    Colonies picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-TT-pLuXR-araC-lasR-TT (X2)
 +
    </li>
 +
</ul>
 +
<br/>
 +
</div>
 +
<div id="page85" class='notebook_page'>
<p>
<p>
-
Innoculation
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; population 1 (oscillator)
+
    August 31, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
Colonies picked
+
    Colony picking
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-lasR-TT-pLasR-LuxI-RBS-YFP
+
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI; 50 colonies, 1 picked
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-araC-LasR-TT
+
    PCR Reaction
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLasR-TetO-RBS-CFP (2 colonies picked)
+
    Add LacO binding site to pLac; 65.2 degrees found to be optimal temperature for elongation
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
Transformation
+
    Transformation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR
+
    pLac-LacO
</p>
</p>
 +
<br/>
<p>
<p>
-
Innoculation
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-LasI-RBS-RFP
+
    August 31
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
Population Testing Prep (pop2, pop3, pop2+pop3)
+
    Glycerol Stock and Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-TT-pLuXR-araC-lasR-TT: Concentration: 115.2
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-TT-pLuXR-araC-lasR-TT: Concentration: 77.7
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion and Gel Extraction
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-TT-pLuXR-araC-lasR-TT (S/P)
 +
    </li>
</div>
</div>
-
<div id="page42" class='notebook_page'><p>
+
<div id="page86" class='notebook_page'>
-
&#9679; 2 ml of medium added to each test tube
+
    <li>
 +
        pLac-LuxR-TT-pLuXR-araC-lasR-TT (S/P)
 +
    </li>
 +
</ul>
 +
<p>
 +
    Colony PCR
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pBad-TetR-TT-pLasR-TetO-CFP (X2)
 +
    </li>
 +
</ul>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Primer 1: I3.24.SP1
 +
    </li>
 +
    <li>
 +
        Primer 2: I3.PSB1A2.R
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; 100 microliters of each population added as necessary
+
    Population Test Results
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        2ml medium, 400ul populations, 1500ul induction
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            pop1+2 (+/-)
 +
        </li>
 +
        <li>
 +
            pop2+3 (+/-)
 +
        </li>
 +
        <li>
 +
            pop1+2+3 (+/-)
 +
        </li>
 +
    </ul>
 +
    <li>
 +
        1ml medium, 200ul populations, 750ul induction
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            pop2+3 (+/-)
 +
        </li>
 +
    </ul>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; .75mL of inducing population added as necessary
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; pop2 w/ w/o inducer
+
    September 1, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; pop3 w/ w/o inducer
+
    Glycerol Stock/Plasmid Isolation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; pop2+pop3 w/ w/o inducer
+
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI
</p>
</p>
 +
</div>
 +
<div id="page87" class='notebook_page'>
 +
<br/>
 +
<br/>
<p>
<p>
-
<strong>August 23</strong>
+
    Colony Picking
</p>
</p>
 +
<br/>
<p>
<p>
-
Glycerol Stock
+
    pLac-LacO
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLasR-TetO-RBS-CFP (X2)
+
    Inoculations
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-lasR-TT-pLasR-LuxI-RBS-YFP
+
    pLac-TetR
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-araC-LasR-TT
+
    pLac-TetR-RBS-RFP
</p>
</p>
 +
<br/>
<p>
<p>
-
Plasmid Isolation
+
    pTet-LacI
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLasR-TetO-RBS-CFP: Concentration: 113.7
+
    pTet-RBS-GFP-T
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLasR-TetO-RBS-CFP: Concentration: 112.0
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-lasR-TT-pLasR-LuxI-RBS-YFP: Concentration: 167.0
+
    September 1
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-araC-LasR-TT: Concentration: 104.2
+
    Ligations
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector1: pLac-LuxR-TT
 +
    </li>
 +
    <li>
 +
        Insert1: pLuxR-araC-LasR-TT
 +
    </li>
 +
    <li>
 +
        Vector2: pLasR
 +
    </li>
 +
    <li>
 +
        Insert2.1: LuxS
 +
    </li>
 +
    <li>
 +
        Insert2.2 araC
 +
    </li>
 +
    <li>
 +
        Vector3: TT
 +
    </li>
</div>
</div>
-
<div id="page43" class='notebook_page'><p>
+
<div id="page88" class='notebook_page'>
-
&#9679; pLac-lasI-RBS-RFP: Concentration: 37.0
+
    <li>
 +
        Insert3: pLsrA-LuxI-RBS-YFP
 +
    </li>
 +
    <li>
 +
        Vector4: pLuxR
 +
    </li>
 +
    <li>
 +
        Insert4: TetR
 +
    </li>
 +
</ul>
 +
<p>
 +
    Transformation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        BBa_C0012: LacI
 +
    </li>
 +
    <li>
 +
        BBa_K091002: LsrK
 +
    </li>
 +
</ul>
<p>
<p>
-
Colonies picked
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
</div>
 +
<div id="page89" class='notebook_page'>
<p>
<p>
-
&#9679; pLac-LuxR
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Population Testing from 8/22 prep after 24 hours
+
    September 2, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; 500 microliters from test tube of populations put in microcentrifuge tubes
+
    Glycerol Stock/Plasmid Isolation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; pop2 w/: slight increase of RFP
+
    pLac-LacO
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9675; pop2 w/o: some RFP
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; pop3 w/: no significant increase of CFP
+
    September 3, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; pop3 w/o: some CFP
+
    Plasmid Isolation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; pop2+pop3 w/: no significant increase of RFP/CFP
+
    pLac-TetR
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; pop2+pop3 w/o: some RFP/CFP
+
    pLac-TetR-RBS-RFP
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Results:
+
    pTet-LacI
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; test tubes were checked after 24 hours
+
    pTet-RBS-GFP-T
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9632;pop2 showed a slight increase in RFP production when it was induced in comparison to its uninduced control
+
    Digestion/Extraction
 +
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    pLac-TetR
</p>
</p>
<p>
<p>
-
&#9632;both pop3 and pop2+pop3 showed no significant differences between its positive and negative controls
+
    pLac-TetR-RBS-RFP
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; after 48 hours
+
    pTet-LacI
</p>
</p>
 +
<br/>
 +
<p>
 +
    pTet-RBS-GFP-T
 +
</p>
 +
<br/>
</div>
</div>
-
<div id="page44" class='notebook_page'><p>
+
<div id="page90" class='notebook_page'>
-
&#9632;pop2 showed increase in RFP production
+
<p>
 +
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9632;pop3 showed no significant increase in CFP production
+
    September 3
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9632;pop2+pop3 showed a sig increase in RFP production but no CFP when it was induced compared to its negative control
+
    Glycerol Stock
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-TT-pLuxR-araC-LasR-TT
 +
    </li>
 +
    <li>
 +
        pBad-TetR-TT-pLas-TetO-CFP
 +
    </li>
 +
</ul>
<p>
<p>
-
Digestion
+
    Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-TT-pLuxR-araC-lasR-TT
 +
    </li>
 +
    <li>
 +
        pBad-TetR-TT-pLas-TetO-CFP
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; plac-LuxR(S/P): Concentration: 19.7
+
    Digestion
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator) (X/P)
 +
    </li>
 +
    <li>
 +
        pBad-TetR-TT-pLas-TetO-CFP (X/P)
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-TT-pLuxR-araC-lasR-TT (S/P)
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pTetR-LasR (E/P): Concentration: gel not used
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pyear-luxS (Inducer pop1)
 +
    </li>
 +
    <li>
 +
        pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
 +
    </li>
 +
</div>
 +
<div id="page91" class='notebook_page'>
 +
 +
    <li>
 +
        pTetR-LuxI (Inducer pop2)
 +
    </li>
 +
    <li>
 +
        pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pLasR-RBS-CFP (X/P): Concentration: gel not used
+
    Colony PCR for I3-98-2 and I4-1-2-
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-TT (S/P): Concentration: 24.0
+
    Digestion
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        TT-pLuxR-LasI-RFP (X/P)
 +
    </li>
 +
    <li>
 +
        pLac-LuxR (S/P)
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; pBad-TetR-TT (S/P): Concentration: 15.4
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LasR-TT-pLasR-LuxI-RBS-YFP (E/P): Concentration: 12.2
+
    September 4, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-araC-lasR-TT (X/P): Concentration: gel not used
+
    Inoculations
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLasR-TetO-RBS-CFP (X/P): Concentration: 5.2
+
    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI (Atc 10 microliters)
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLasR-TetO-RBS-CFP (X/P): Concentration: 5.2
+
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI (IPTG 10 microliters)
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-LasI-RBS-RFP (X/P): Concentration: gel not used
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Innoculation
+
    September 4
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pyear-luxS
+
    Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pBad-TetR-TT-pLasR-TetO-CFP: Concentration: 190.4
 +
    </li>
 +
</ul>
</div>
</div>
-
<div id="page45" class='notebook_page'><p>
+
<div id="page92" class='notebook_page'>
-
&#9679; pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
+
-
</p>
+
<p>
<p>
-
&#9679; pTetR-LuxI
+
    Digestion
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pBad-TetR-TT-pLasR-TetO-CFP (X/P): Concentration: 20.5
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
+
    Colonies picked from I4-1-2
</p>
</p>
 +
<br/>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLac-lasI
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
+
    September 5, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-lasI-RBS-RFP
+
    Inoculations
 +
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI
</p>
</p>
<p>
<p>
-
Ligation
+
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector: pBad-TetR-TT
+
    1 with Atc
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert: plasR-TetO-RBS-CFP
+
    1 with IPTG
</p>
</p>
 +
<br/>
<p>
<p>
-
<strong>August 24</strong>
+
    1 with No induction
</p>
</p>
 +
<br/>
<p>
<p>
-
Plasmid Isolation
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pluxR-LasI-RBS-RFP: Concentration: 42.6
+
    September 5
</p>
</p>
 +
<br/>
<p>
<p>
-
Digestion
+
    Glycerol Stock and Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLasR-LuxS
 +
    </li>
 +
    <li>
 +
        pLasR-araC
 +
    </li>
 +
    <li>
 +
        TT-pLsrA-LuxI-RBS-YFP
 +
    </li>
 +
</div>
 +
<div id="page93" class='notebook_page'>
 +
    <li>
 +
        pLuxR-TetR
 +
    </li>
 +
    <li>
 +
        LacI
 +
    </li>
 +
    <li>
 +
        LsrK
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pLuxR-LasI-RBS-RFP (X/P)
+
    Digestion and Gel Extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLasR-LuxS
 +
    </li>
 +
    <li>
 +
        pLasR-araC
 +
    </li>
 +
    <li>
 +
        TT-pLsrA-LuxI-RBS-YFP
 +
    </li>
 +
    <li>
 +
        pLuxR-TetR
 +
    </li>
 +
    <li>
 +
        LacI
 +
    </li>
 +
    <li>
 +
        LsrK
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pLuxR-araC-lasR-TT (X/P)
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac
 +
    </li>
 +
    <li>
 +
        LasI-RFP
 +
    </li>
 +
</ul>
 +
<br/>
</div>
</div>
-
<div id="page46" class='notebook_page'><p>
+
<div id="page94" class='notebook_page'>
-
Innoculation
+
<p>
 +
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-LasI-RBS-RFP
+
    September 6, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-araC-LasR-TT
+
    Sequencing
</p>
</p>
 +
<br/>
 +
<p>
<p>
-
&#9679; pLsrA-LuxI-YFP
+
    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI
</p>
</p>
 +
<br/>
<p>
<p>
-
August 25
+
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI
</p>
</p>
 +
<br/>
<p>
<p>
-
Glycerol Stock
+
    pLac-LacO
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; plac-LuxR
+
    RFP-LVA
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pBad-TetR-TT-plasR-TetO-RBS-CFP
+
    Inoculations
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pBad-TetR-TT-plasR-TetO-RBS-CFP
+
    Chlorabackbone
 +
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI
</p>
</p>
<p>
<p>
-
&#9679; pLuxR-araC-lasR-TT
+
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI
</p>
</p>
 +
<br/>
<p>
<p>
-
Plasmid Isolation
+
    1 with Atc
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-LasI-RBS-RFP: Concentration: 89.1
+
    1 with IPTG
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-araC-LasR-TT: Concentration: 152.9
+
    1 with No induction
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pBad-TetR-TT-plasR-TetO-RBS-CFP: Concentration: 200.0
+
    Digestion/Extraction
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pBad-TetR-TT-plasR-TetO-RBS-CFP: Concentration: 156.0
+
    Chlorabackbone
</p>
</p>
 +
<br/>
 +
<br/>
</div>
</div>
-
<div id="page47" class='notebook_page'><p>
+
<div id="page95" class='notebook_page'>
-
Digestion and Gel Extraction
+
<p>
 +
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-LasI-RBS-RFP: (X/P): Concentration: 9.5
+
    September 6
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-araC-LasR-TT: (X/P): gel not used
+
    Plasmid Isolation and Glycerol Stock
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac
 +
    </li>
 +
    <li>
 +
        LasI-RFP
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-TT-pLuxR-araC-LasR-TT
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pBad-TetR-TT-plasR-TetO-RBS-CFP: (X/P): gel not used
+
    Digestion and Gel Extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac
 +
    </li>
 +
    <li>
 +
        LasI-RFP
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-TT-pLuxR-araC-LasR-TT
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pBad-TetR-TT-plasR-TetO-RBS-CFP: (X/P): gel not used
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector1:pLac-LuxR-TT-pLuxR-araC-LasR-TT
 +
    </li>
 +
    <li>
 +
        Insert1: pBad-&gt;&gt;CFP
 +
    </li>
 +
    <li>
 +
        Vector2: pLsrA
 +
    </li>
 +
    <li>
 +
        Insert2: LacI
 +
    </li>
 +
    <li>
 +
        Vector3: pLasR-LuxS
 +
    </li>
 +
</div>
 +
<div id="page96" class='notebook_page'>
 +
    <li>
 +
        Insert3: RBS-CFP
 +
    </li>
 +
    <li>
 +
        Vector4: Term-Term
 +
    </li>
 +
    <li>
 +
        Insert4: pLuxR-TetR
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
Ligation
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector1: pLac-LuxR-TT
+
    September 7, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert1: pLuxR-LasI-RBS-RFP
+
    Inoculation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector2: pTetR-LasR
+
    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert2: pLasR-CFP
+
    pLac-RBS-GFP-T
</p>
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    pTet-RBS-GFP-T-pLac-TetR
 +
</p>
 +
<p dir="ltr">
 +
    pLac-RBS-GFP-T-pTet-LacI
 +
</p>
 +
<p dir="ltr">
 +
    RFP-LVA
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LacO
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI
 +
</p>
 +
<p dir="ltr">
 +
    pTet-LasI
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LuxR-DT-pLux-TetR
 +
</p>
 +
<p dir="ltr">
 +
    LuxR-DT-pLux-TetR
 +
</p>
 +
<p dir="ltr">
 +
    LuxR-DT
 +
</p>
 +
</div>
 +
<div id="page97" class='notebook_page'>
 +
<p dir="ltr">
 +
    pLux-TetR
 +
</p>
 +
<p dir="ltr">
 +
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
<p dir="ltr">
 +
    pLas-LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
<p dir="ltr">
 +
    LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
<p dir="ltr">
 +
    LacI-RFP-ST
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LuxI
 +
</p>
 +
<p dir="ltr">
 +
    pTet-LasR-DT
 +
</p>
 +
<p dir="ltr">
 +
    LasR-DT
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LacO
 +
</p>
 +
<p dir="ltr">
 +
    pTet-LacI
 +
</p>
 +
<p dir="ltr">
 +
    RBS-GFP-ST
 +
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector3: pLac-LuxR-TT
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert3: pLuxR-LasI-RBS-RFP
+
    September 7
</p>
</p>
 +
<br/>
<p>
<p>
-
Primer Dilution
+
    Test Digestions
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac
 +
    </li>
 +
    <li>
 +
        LasI-RFP
 +
    </li>
 +
</div>
 +
<div id="page98" class='notebook_page'>
 +
    <li>
 +
        pLac-LuxR-TT-pLuxR-araC-LasR-TT
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; I3.PsB1A2.R +661ul NFH2O -&gt; 1:10 Dulition
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Term-Term
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; I3.DT.F +634ul NFH2O -&gt; 1:10 Dulition
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; I3.FT.M +700ul NFH2O -&gt; 1:10 Dulition
+
    September 8, 2012
 +
</p>
 +
<br/>
 +
<p>
 +
    Time-Lapse Experiment to observe toggle switch
 +
</p>
 +
<br/>
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP
 +
</p>
 +
<p>
 +
    pLac-RBS-GFP-T
 +
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    pTet-RBS-GFP-T-pLac-TetR
 +
</p>
 +
<p dir="ltr">
 +
    pLac-RBS-GFP-T-pTet-LacI
 +
</p>
 +
<p dir="ltr">
 +
    RFP-LVA
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LacO
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI
 +
</p>
 +
<p dir="ltr">
 +
    pTet-LasI
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LuxR-DT-pLux-TetR
</p>
</p>
</div>
</div>
-
<div id="page48" class='notebook_page'><p>
+
<div id="page99" class='notebook_page'>
-
Colony PCR
+
<p dir="ltr">
 +
    LuxR-DT-pLux-TetR
</p>
</p>
 +
<p dir="ltr">
 +
    LuxR-DT
 +
</p>
 +
<p dir="ltr">
 +
    pLux-TetR
 +
</p>
 +
<p dir="ltr">
 +
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
<p dir="ltr">
 +
    pLas-LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
<p dir="ltr">
 +
    LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
<p dir="ltr">
 +
    LacI-RFP-ST
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LuxI
 +
</p>
 +
<p dir="ltr">
 +
    pTet-LasR-DT
 +
</p>
 +
<p dir="ltr">
 +
    LasR-DT
 +
</p>
 +
<p dir="ltr">
 +
    pTet-LacI
 +
</p>
 +
<p dir="ltr">
 +
    GFP
 +
</p>
 +
<br/>
<p>
<p>
-
&#9679; pBad-TetR-TT-plasR-TetO-RBS-CFP
+
    Ligations
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-araC-LasR-TT
+
    pLac-TetR into Chlora
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; Primer 1: I3.24.SP1
+
    pLac-RBS-GFP-T into Chlora
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; Primer 2: I3.PSB1A2.R
+
    pTet-RBS-GFP-T into Chlora
</p>
</p>
 +
<br/>
<p>
<p>
-
Innoculation
+
    pTet-LacI into Chlora
</p>
</p>
 +
<br/>
 +
<br/>]
 +
</div>
 +
<div id="page100" class='notebook_page'>
<p>
<p>
-
&#9679; pyear-luxS
+
    Digestion/Extraction
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
+
    GFP
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-LuxI
+
    pLac-LacO
 +
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP
</p>
</p>
<p>
<p>
-
&#9679; pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
+
    pLac-RBS-GFP-T
 +
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    pTet-RBS-GFP-T-pLac-TetR
 +
</p>
 +
<p dir="ltr">
 +
    pLac-RBS-GFP-T-pTet-LacI
 +
</p>
 +
<p dir="ltr">
 +
    RFP-LVA
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LacO
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI
 +
</p>
 +
<p dir="ltr">
 +
    pTet-LasI
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LuxR-DT-pLux-TetR
 +
</p>
 +
<p dir="ltr">
 +
    LuxR-DT-pLux-TetR
 +
</p>
 +
<p dir="ltr">
 +
    LuxR-DT
 +
</p>
 +
<p dir="ltr">
 +
    pLux-TetR
 +
</p>
 +
<p dir="ltr">
 +
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
</div>
 +
<div id="page101" class='notebook_page'>
 +
<p dir="ltr">
 +
    pLas-LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
<p dir="ltr">
 +
    LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
<p dir="ltr">
 +
    LacI-RFP-ST
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LuxI
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-lasI
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
+
    September 8
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-lasI-RBS-RFP
+
    Colonies Picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        14-7-5
 +
    </li>
 +
    <li>
 +
        14-7-7
 +
    </li>
 +
    <li>
 +
        14-8-2
 +
    </li>
 +
</ul>
 +
<br/>
</div>
</div>
-
<div id="page49" class='notebook_page'><p>
+
<div id="page102" class='notebook_page'>
-
<strong>August 26</strong>
+
 
 +
<p>
 +
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Colonies Picked
+
    September 9, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-araC-LasR-TT: 1 colony picked
+
    Colony Picking
</p>
</p>
 +
<br/>
<p>
<p>
-
Population testing
+
    pLac-TetR, 1 colony picked
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; 2ml broth, 1ml induction, 100ul population, 30 hour incubation
+
    pLac-RBS-GFP-T, 1 colony picked
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; pop1+2 (+/-): no fluorescence seen
+
    pTet-LacI, 1 colony picked
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9675; pop2+3 (+/-): no fluorescence seen
+
    Ligation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; pop1+2+3 (+/-): no fluorescence seen
+
    pLac-LacO-GFP
</p>
</p>
 +
<br/>
 +
<br/>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
<br/>
<p>
<p>
-
<strong>August 27</strong>
+
    September 9
</p>
</p>
 +
<br/>
<p>
<p>
-
Plasmid Isolation
+
    Glycerol Stock and Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-TT-pLuxR-araC-LasR-TT-pBad-TetR-TT-pLasR-TetO-CFP
 +
    </li>
 +
    <li>
 +
        pLsrA-LacI
 +
    </li>
 +
    <li>
 +
        pLasR-LuxS-RBS-CFP
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; pLuxR-araC-LaR-TT: Concentration: 164.3
+
    Digestion and Gel Extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-TT-pLuxR-araC-LasR-TT-pBad-TetR-TT-pLasR-TetO-CFP
 +
    </li>
 +
</div>
 +
<div id="page103" class='notebook_page'>
 +
    <li>
 +
        pLsrA-LacI
 +
    </li>
 +
    <li>
 +
        pLasR-LuxS-RBS-CFP
 +
    </li>
 +
</ul>
<p>
<p>
-
Digestion and Gel extraction
+
    Ligations
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector1: Term-Term
 +
    </li>
 +
    <li>
 +
        Insert1-1: pLsrA-LacI
 +
    </li>
 +
    <li>
 +
        Insert1-2: pLuxR-TetR
 +
    </li>
 +
    <li>
 +
        Insert1-3: pLsrA-LuxI-RBS-YFP
 +
    </li>
 +
    <li>
 +
        Insert1-4: pLasR-LuxS-RBS-CFP
 +
    </li>
 +
    <li>
 +
        Insert1-5: pLasR-araC
 +
    </li>
 +
    <li>
 +
        Vector2: pLac
 +
    </li>
 +
    <li>
 +
        Insert2: 13-12-8
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-araC-LaR-TT (X/P): Concentration: 55.1
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Innoculation
+
    September 10, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pyear-luxS
+
    Glycerol Stock/Miniprep
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
+
    pLac-TetR
</p>
</p>
 +
<br/>
 +
<p>
 +
    pLac-RBS-GFP-T
 +
</p>
 +
<br
</div>
</div>
-
<div id="page50" class='notebook_page'><p>
+
<div id="page104" class='notebook_page'>
-
&#9679; pTetR-LuxI
+
<p>
 +
    pTet-LacI
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
+
    Gel Extraction
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-lasI
+
    pLac-LacO-GFP
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
+
    pTet-LacI
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; pLuxR-lasI-RBS-RFP
+
    Ligation
</p>
</p>
 +
<br/>
<p>
<p>
-
Ligation
+
    pTet-LacI into pLac-LacO-RBS-GFP-ST
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector1: pBad-tetR-TT
+
    pLac-RBS-GFP-T-pTet-LacI into chlora backbone
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; Insert1: pLasR-TetO-RBS-CFP
+
    Triple Population and AND Gate Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 0:1 20 colonies seen
+
    September 10
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 2:1 50 colonies seen
+
    Colonies Picked
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        I4-11-2
 +
    </li>
 +
    <li>
 +
        I4-11-3
 +
    </li>
 +
    <li>
 +
        I4-11-4
 +
    </li>
 +
    <li>
 +
        I4-11-5
 +
    </li>
 +
    <li>
 +
        I4-11-6
 +
    </li>
 +
    <li>
 +
        I4-8-4
 +
    </li>
 +
</div>
 +
<div id="page105" class='notebook_page'>
 +
    <li>
 +
        I4-8-5
 +
    </li>
 +
    <li>
 +
        I4-8-6
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; Vector2:pLac-LuxR-TT
+
    Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pLac
 +
    </li>
 +
    <li>
 +
        LuxR
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9679; Insert2: pLuXR-araC-lasR-TT
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 0:1 5 colonies seen
+
    September 11, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 2:1 25 colonies seen
+
    PCR
</p>
</p>
 +
<br/>
 +
<p>
 +
    pLac-LacO
 +
</p>
 +
<br/>
 +
<p>
 +
    pLac-LacO x2
 +
</p>
 +
<br/>
 +
<br/>
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
<br/>
 +
<p>
 +
    September 12, 2012
 +
</p>
 +
<br/>
 +
<p>
 +
    Digestion/Gel Extraction
 +
</p>
 +
<br/>
 +
<p>
 +
    pTet-LacI-pLac-LacO-RBS-GFP-ST
 +
</p>
 +
<br/>
 +
<br/>
</div>
</div>
-
<div id="page51" class='notebook_page'><p>
+
<div id="page106" class='notebook_page'>
-
<strong>August 28</strong>
+
<p>
 +
    Experiments
</p>
</p>
 +
<br/>
<p>
<p>
-
Colony PCR
+
    Toggle with 20 micro liters IPTG
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pBad-tetR-TT-pLasR-TetO-RBS-CFP
+
    Toggle with 40 micro liters ATc
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-TT- pLuXR-araC-lasR-TT
+
    Strain 1 of dual population toggle with inducer strain
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; Primer 1: I3.24.SP1
+
    Strain 2 of dual population toggle with inducer strain
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9675; Primer 2: I3.PSB1A2.R
+
    Ligation
</p>
</p>
 +
<br/>
<p>
<p>
-
Population Testing
+
    pLac-LacO-RBS-GFP-ST-pTet-LacI into chlora backbone
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
&#9679; 200ul medium, 200ul induction, 4ul population, 24 hour incubation
+
    Triple Population and AND Gate
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; pop1+2 (-): no fluorescence seen
+
    September 13
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; pop1+2 (+): some sign of RFP/YFP
+
    Glycerol Stock and Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        I4-8-4
 +
    </li>
 +
    <li>
 +
        I4-8-5
 +
    </li>
 +
    <li>
 +
        I4-8-6
 +
    </li>
 +
    <li>
 +
        I4-11-2
 +
    </li>
 +
    <li>
 +
        I4-11-3
 +
    </li>
 +
    <li>
 +
        I4-11-4
 +
    </li>
 +
    <li>
 +
        I4-11-5
 +
    </li>
 +
</div>
 +
<div id="page107" class='notebook_page'>
 +
    <li>
 +
        I4-11-6
 +
    </li>
 +
 +
    <li>
 +
        I4-13-1
 +
    </li>
 +
    <li>
 +
        I4-13-2
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9675; pop2+3 (-): no fluorescence seen
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pyear-luxS
 +
    </li>
 +
    <li>
 +
        pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
 +
    </li>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
    <li>
 +
        pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
 +
    </li>
 +
    <li>
 +
        pLac-lasI
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9675; pop2+3 (+): no fluorescence seen
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector: Const. Promoter
 +
    </li>
 +
    <li>
 +
        Insert1-1: I3-28-4
 +
    </li>
 +
    <li>
 +
        Insert1-2: I4-6-6
 +
    </li>
 +
    <li>
 +
        Insert1-3: I4-16-2
 +
    </li>
 +
    <li>
 +
        Vector2: TT-pLasR-LuxS-RBS-CFP
 +
    </li>
 +
</div>
 +
<div id="page108" class='notebook_page'>
 +
    <li>
 +
        Insert2: TT-pLuxR-TetR
 +
    </li>
 +
    <li>
 +
        Vector3: pLac
 +
    </li>
 +
    <li>
 +
        Insert3: LuxR
 +
    </li>
 +
    <li>
 +
        Vector4: pLuxR
 +
    </li>
 +
    <li>
 +
        Insert4: LasI-RFP
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9675; pop1+2+3 (-): no fluorescence seen
+
    Ligations for Chlora
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        I4-14-7
 +
    </li>
 +
    <li>
 +
        I4-14-8
 +
    </li>
 +
    <li>
 +
        I4-14-10
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9675; pop1+2+3 (+): no fluorescence seen
+
    Transformation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        I4-18-1
 +
    </li>
 +
    <li>
 +
        I4-18-2
 +
    </li>
 +
    <li>
 +
        I4-18-3
 +
    </li>
 +
    <li>
 +
        I401804
 +
    </li>
 +
</ul>
 +
<p>
 +
    Innoculation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        I3-28-2
 +
    </li>
</div>
</div>
-
<div id="page52" class='notebook_page'><p>
+
<div id="page109" class='notebook_page'>
-
Ligation
+
    <li>
 +
        I3-30-4
 +
    </li>
 +
    <li>
 +
        I3-11-1
 +
    </li>
 +
    <li>
 +
        I3-46-4
 +
    </li>
 +
</ul>
 +
<br/>
 +
<p>
 +
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector:pLac-LuxR-TT
+
    September 14, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert: pLuXR-araC-lasR-TT
+
    Ligation
 +
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP into Chlora backbone
</p>
</p>
<p>
<p>
-
&#9675; 0:1 0 colonies seen
+
    pLac-RBS-GFP-T into Chlora backbone
</p>
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    pTet-RBS-GFP-T-pLac-TetR into Chlora backbone
 +
</p>
 +
<p dir="ltr">
 +
    pLac-RBS-GFP-T-pTet-LacI into Chlora backbone
 +
</p>
 +
<p dir="ltr">
 +
    RFP-LVA into Chlora backbone
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LacO into Chlora backbone
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI into Chlora backbone
 +
</p>
 +
<p dir="ltr">
 +
    pTet-LasI into Chlora backbone
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LuxR-DT-pLux-TetR into Chlora backbone
 +
</p>
 +
<p dir="ltr">
 +
    LuxR-DT-pLux-TetR into Chlora backbone
 +
</p>
 +
</div>
 +
<div id="page110" class='notebook_page'>
 +
 +
<p dir="ltr">
 +
    LuxR-DT into Chlora backbone
 +
</p>
 +
<p dir="ltr">
 +
    pLux-TetR into Chlora backbone
 +
</p>
 +
<p dir="ltr">
 +
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI into Chlora backbone
 +
</p>
 +
<p dir="ltr">
 +
    pLas-LacI-RFP-ST-pLac-LuxI into Chlora backbone
 +
</p>
 +
<p dir="ltr">
 +
    LacI-RFP-ST-pLac-LuxI into Chlora backbone
 +
</p>
 +
<p dir="ltr">
 +
    LacI-RFP-ST into Chlora backbone
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LuxI into Chlora backbone
 +
</p>
 +
<p dir="ltr">
 +
    pLac-LacO x2-RBS-GFP-ST-pTet-LacI
 +
</p>
 +
<br/>
<p>
<p>
-
&#9675; 2:1 30 colonies seen
+
    Glycerol Stock/Plasmid Isolation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 4:1 15 colonies seen
+
    pLac-LacO-GFP
</p>
</p>
 +
<br/>
 +
<br/>
<p>
<p>
-
Digestion and Gel Extraction
+
    Digestion
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pBad-TetR-TT (S/P): Concentration: 88.3
+
    pLac-LacO-GFP
</p>
</p>
 +
<br/>
<p>
<p>
-
Innoculation
+
    pTet-LacI
</p>
</p>
 +
<br/>
 +
<p dir="ltr">
 +
    pLac-LacO x2-RBS-GFP-ST-pTet-LacI
 +
</p>
 +
<br/>
<p>
<p>
-
&#9679; pyear-luxS
+
    Inoculation
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
+
    pLac-GFP-pTet-LacI
</p>
</p>
 +
<br/>
 +
<br/>
 +
</div>
 +
<div id="page111" class='notebook_page'>
<p>
<p>
-
&#9679; pTetR-LuxI
+
    Triple Population and AND Gate
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
+
    September 15
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
+
    Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        I4-16-4 CP-LasR
 +
    </li>
 +
    <li>
 +
        I4-16-5 CP-LsrK
 +
    </li>
 +
    <li>
 +
        I4-16-6 CP-LuxR
 +
    </li>
 +
    <li>
 +
        I4-17-4 TT-pLasR-&gt;&gt;TetR
 +
    </li>
 +
    <li>
 +
        I4-17-6 pLac-LuxR
 +
    </li>
 +
    <li>
 +
        I4-17-8 pLuxR-LasI-RFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Colonies picked
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        I4-19-19
 +
    </li>
 +
    <li>
 +
        I4-22-8
 +
    </li>
 +
    <li>
 +
        I4-22-9
 +
    </li>
 +
    <li>
 +
        I4-21-2
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        I4-16-4
 +
    </li>
</div>
</div>
-
<div id="page53" class='notebook_page'><p>
+
<div id="page112" class='notebook_page'>
-
<strong>August 29</strong>
+
    <li>
 +
        I4-16-5
 +
    </li>
 +
    <li>
 +
        I4-16-6
 +
    </li>
 +
    <li>
 +
        I4-17-4
 +
    </li>
 +
    <li>
 +
        I4-17-6
 +
    </li>
 +
    <li>
 +
        I4-17-8
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligations
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector1: pTetR-LasR
 +
    </li>
 +
    <li>
 +
        Insert1: TT-pLasR-LuxS-RFP-TT-pLuxR-TetR
 +
    </li>
 +
    <li>
 +
        Vector2: Term-Term
 +
    </li>
 +
    <li>
 +
        Insert2: pLac-LuxR
 +
    </li>
 +
    <li>
 +
        Vector3: pLuxR-LasII-RFP
 +
    </li>
 +
    <li>
 +
        Insert3: TT-pLsrA-LacI
 +
    </li>
 +
</ul>
<p>
<p>
-
Population Test Prep
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pyear-luxS
 +
    </li>
 +
    <li>
 +
        pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
 +
    </li>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
</div>
 +
<div id="page113" class='notebook_page'>
 +
    <li>
 +
        pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
 +
    </li>
 +
    <li>
 +
        pLac-lasI
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
<br/>
 +
</div>
 +
<div id="page114" class='notebook_page'>
<p>
<p>
-
&#9679; 2ml medium, 400ul populations, 1500ul induction
+
    Triple Population and AND Gate
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; pop1+2 (+/-)
+
    September 18
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; pop2+3 (+/-)
+
    Glycerol Stock and Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pTetR-LasR-TT-pLasR-LuxS-CFP-TT-pLuxR-TetR
 +
    </li>
 +
    <li>
 +
        TT-pLac-LuxR
 +
    </li>
 +
    <li>
 +
        pLuxR-LasI-RFP-TT-pLsrA-LacI
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9675; pop1+2+3 (+/-)
+
    Digestion and Gel Extraction
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        I4-10-7
 +
    </li>
 +
    <li>
 +
        I4-25-14
 +
    </li>
 +
    <li>
 +
        I4-25-16
 +
    </li>
 +
    <li>
 +
        I4-25-18
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; 1ml medium, 200ul populations, 750ul induction
+
    Ligation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector1: CP-LasR
 +
    </li>
 +
    <li>
 +
        Insert1: TT
 +
    </li>
 +
    <li>
 +
        Vector2: TT-pLsrA-LuxI-YFP
 +
    </li>
 +
    <li>
 +
        Insert 2: TT-pLasR-araC
 +
    </li>
 +
</div>
 +
<div id="page115" class='notebook_page'>
 +
    <li>
 +
        Vector3: TT
 +
    </li>
 +
    <li>
 +
        Insert3.1: pTetR-&gt;&gt;TetR
 +
    </li>
 +
    <li>
 +
        Insert3.2: pLuxR-&gt;&gt;LacI
 +
    </li>
 +
    <li>
 +
        Vector4: CP-LsrK
 +
    </li>
 +
    <li>
 +
        Insert4: TT-pLac-LuxR
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9675; pop2+3 (+/-)
+
    Innoculation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Pop3 AND Gate
 +
    </li>
 +
    <li>
 +
        Pop2 AND Gate
 +
    </li>
 +
    <li>
 +
        Pop1 AND Gate
 +
    </li>
 +
</ul>
<p>
<p>
-
<strong>August 30</strong>
+
    Testing AND Gate
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Pop2 and 3
 +
    </li>
 +
    <li>
 +
        Pop1 and 3
 +
    </li>
 +
    <li>
 +
        Pop 1 2 3
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
Ligation
+
    Triple Population and AND Gate
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Vector: pBad-TetR-TT
+
    September 19
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; Insert: plasR-TetO-CFP
+
    Plate Experiments for AND Gate
</p>
</p>
 +
<br/>
 +
<br/>
 +
<br/>
 +
<br/>
 +
<br/>
 +
<br/>
 +
</div>
 +
<div id="page116" class='notebook_page'>
<p>
<p>
-
&#9675; 0:1 100 Colonies seen
+
    Triple Population and AND Gate
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 1:1 200 Colonies seen
+
    September 20
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; 4:1 200 Colonies seen
+
    Glycerol Stock and Plasmid Isolation
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Cp-LasR-TT
 +
    </li>
 +
    <li>
 +
        TT-pLsrA-&gt;&gt;araC
 +
    </li>
 +
    <li>
 +
        TT-pTetR-&gt;&gt;TetR
 +
    </li>
 +
    <li>
 +
        CP-LsrK-&gt;&gt;LuxR
 +
    </li>
 +
    <li>
 +
        TT-pLuxR-&gt;&gt;LacI
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestions (repeated 3X)
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Cp-LasR-TT
 +
    </li>
 +
    <li>
 +
        TT-pLsrA-&gt;&gt;araC
 +
    </li>
 +
    <li>
 +
        TT-pTetR-&gt;&gt;TetR
 +
    </li>
 +
    <li>
 +
        CP-LsrK-&gt;&gt;LuxR
 +
    </li>
 +
    <li>
 +
        TT-pLuxR-&gt;&gt;LacI
 +
    </li>
 +
</ul>
 +
<br/>
</div>
</div>
-
<div id="page54" class='notebook_page'><p>
+
<div id="page117" class='notebook_page'>
-
Colony PCR
+
<p>
 +
    Triple Population and AND Gate
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-TT-pLuXR-araC-lasR-TT (X2)
+
    September 22
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9675; Primer 1: I3.24.SP1
+
    Ligations (Repeated)
</p>
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector1: CP-LasR
 +
    </li>
 +
    <li>
 +
        Insert1: TT
 +
    </li>
 +
    <li>
 +
        Vector2: TT-pLsrA-LuxI-YFP
 +
    </li>
 +
    <li>
 +
        Insert 2: TT-pLasR-araC
 +
    </li>
 +
    <li>
 +
        Vector3: TT
 +
    </li>
 +
    <li>
 +
        Insert3.1: pTetR-&gt;&gt;TetR
 +
    </li>
 +
    <li>
 +
        Insert3.2: pLuxR-&gt;&gt;LacI
 +
    </li>
 +
    <li>
 +
        Vector4: CP-LsrK
 +
    </li>
 +
    <li>
 +
        Insert4: TT-pLac-LuxR
 +
    </li>
 +
</ul>
 +
<br/>
<p>
<p>
-
&#9675; Primer 2: I3.PSB1A2.R
+
    Single and Double Population Projects
</p>
</p>
 +
<br/>
<p>
<p>
-
Colonies picked
+
    September 22, 2012
</p>
</p>
 +
<br/>
<p>
<p>
-
&#9679; pLac-LuxR-TT-pLuXR-araC-lasR-TT (X2)
+
    Single toggle experiment - No induction at start, check every 6 hours, induce with IPTG after first 6 hours, and induce with ATc in new tube after 12 Hours
</p>
</p>
 +
<br/>
</div>
</div>
-
 
+
<div id="page118" class='notebook_page'>
 +
<p>
 +
    Triple Population and AND Gate
 +
</p>
 +
<br/>
 +
<p>
 +
    September 22
 +
</p>
 +
<br/>
 +
<p>
 +
    Ligations (Repeated)
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Vector1: CP-LasR
 +
    </li>
 +
    <li>
 +
        Insert1: TT
 +
    </li>
 +
    <li>
 +
        Vector2: TT-pLsrA-LuxI-YFP
 +
    </li>
 +
    <li>
 +
        Insert 2: TT-pLasR-araC
 +
    </li>
 +
    <li>
 +
        Vector3: TT
 +
    </li>
 +
    <li>
 +
        Insert3.1: pTetR-&gt;&gt;TetR
 +
    </li>
 +
    <li>
 +
        Insert3.2: pLuxR-&gt;&gt;LacI
 +
    </li>
 +
    <li>
 +
        Vector4: CP-LsrK
 +
    </li>
 +
    <li>
 +
        Insert4: TT-pLac-LuxR
 +
    </li>
 +
</ul>
 +
<br/>
 +
</div>
 +
<div id="page119" class='notebook_page'>
 +
<p>
 +
    Triple Population and AND Gate
 +
</p>
 +
<br/>
 +
<p>
 +
    September 23
 +
</p>
 +
<br/>
 +
<p>
 +
    Digestions
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Cp-LasR-TT
 +
    </li>
 +
    <li>
 +
        TT-pLsrA-&gt;&gt;araC
 +
    </li>
 +
    <li>
 +
        TT-pTetR-&gt;&gt;TetR
 +
    </li>
 +
    <li>
 +
        CP-LsrK-&gt;&gt;LuxR
 +
    </li>
 +
    <li>
 +
        TT-pLuxR-&gt;&gt;LacI
 +
    </li>
 +
</ul>
 +
<p>
 +
    Innoculations for AND Gate
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Pop1
 +
    </li>
 +
    <li>
 +
        Pop2
 +
    </li>
 +
    <li>
 +
        Pop3
 +
    </li>
 +
</ul>
 +
<p>
 +
    Testing AND Gate
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        Pop 1 3
 +
    </li>
 +
    <li>
 +
        Pop 2 3
 +
    </li>
 +
    <li>
 +
        Pop 1 2 3
 +
    </li>
 +
</ul>
 +
</div>
 +
<div id="page120" class='notebook_page'>
 +
<p>
 +
    Innoculation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        pyear-luxS
 +
    </li>
 +
    <li>
 +
        pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
 +
    </li>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
    <li>
 +
        pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
 +
    </li>
 +
    <li>
 +
        pLac-lasI
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
 +
    </li>
 +
</ul>
 +
<br/>
 +
<br/>
 +
</div>
 +
<div id="page121" class='notebook_page'>
 +
<p>
 +
    Triple Population and AND Gate
 +
</p>
 +
<br/>
 +
<p>
 +
    September 23
 +
</p>
 +
<br/>
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        TT-pLsrA-LuxI-&gt;&gt;araC
 +
    </li>
 +
    <li>
 +
        TT-pTetR-TetR
 +
    </li>
 +
    <li>
 +
        CP-LsrK-&gt;&gt;LuxR
 +
    </li>
 +
    <li>
 +
        TT-pLuxR-LacI
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestions and Gel Extraction
 +
</p>
 +
<br/>
 +
<ul>
 +
    <li>
 +
        TT-pLsrA-LuxI-&gt;&gt;araC
 +
    </li>
 +
    <li>
 +
        TT-pTetR-TetR
 +
    </li>
 +
    <li>
 +
        CP-LsrK-&gt;&gt;LuxR
 +
    </li>
 +
    <li>
 +
        TT-pLuxR-LacI
 +
    </li>
 +
</ul>
 +
<br/>
</html>
</html>

Revision as of 01:09, 4 October 2012

Cover page

June 6


Single and Double Population Projects


Transformation


  • BBa_J61002: “RFP” promoter
  • BBa_K081012: GFP protein generator (RBS-GFP)
  • BBa_J23103: constitutive family member
  • Bba_J04450: RFP Coding Device (pLas-RBS-RFP-Term)

Single and Double Population Projects


June 7


Colonies picked and glycerol stocked made


  • RFP promoter: 5 colonies seen: 1 colony picked
  • RBS-GFP: 2 colonies seen: 1 colony picked
  • constitutive family promoter: 2 colonies seen: 1 colony picked
  • pLas-RBS-RFP-Term: 6 colonies seen: 1 colony picked

Single and Double Population Projects


June 8


Plasmid isolation


  • RBS-GFP
    • Concentration:111.2 ng/ul
  • constututive family member
    • Concentration: 109.8 ng/ul
  • pLasI-RBS-RFP-Term
    • Concentration: 113.9 ng/ul

Single and Double Population Projects


June 11


Digestion


  • RBS-GFP (E/S)---(wrong enzymes used)
  • constitutive promoter (E/X)---(wrong enzymes used)
  • constitutive promoter (S/P)
  • pLasI-RBS-RFP-Term (X/P)

Single and Double Population Projects


June 12


Digestion


  • RBS-GFP (X/P)

Ligation


  • Vector: constitutive promoter
  • Insert: RBS-GFP
  • 0:1--- 0 colonies counted
  • 2:1--- 7 colonies counted

June 13


Incubate constitutive promoter-RBS-GFP in LB carb broth overnight @ 37*C and 220 rpm


  • 2:1--- 2 colonies picked

Single and Double Population Projects


June 14


Glycerol stock and miniprep


  • constitutive promoter-RBS-GFP
    • colony 1 picked: 91 ng/ul
    • colony 2 picked: 97 ng/ul

Test Digestion for each plasmid: correct results received



Single and Double Population Projects


June 15


Plasmid isolation


  • prom-RBS-GFP: Concentration: 127 ng/μl

Single and Double Population Projects


July 16


Transformations


  • Bba_I763035 pTetR-RBS-LacI-RBS-GFP-Term-Term
  • Bba_K093005 RBS-RFP
  • Bba_R0010 pLacI
  • Bba_C0040 TetR

Single and Double Population Projects


July 17


  • Bba_I763035 pTetR-RBS-LacI-RBS-GFP-Term-Term
    • 2 colonies picked
  • Bba_K093005 RBS-RFP
    • 2 colonies picked
  • Bba_R0010 pLacI
    • 2 colonies picked
  • Bba_C0040 TetR
    • 2 colonies picked

Single and Double Population Projects


July 18


Glycerol stocks made


  • pTetR-RBS-LacI-RBS-GFP-Term-Term
    • Concentration: 236.2 ng/μl
  • RBS-RFP
    • Concentration: 92.9 ng/μl
  • pLacI
    • Concentration: 60.6 ng/μl
  • TetR
    • Concentration: 110.9 ng/μl

Digestion


  • pTetR-RBS-LacI-RBS-GFP-Term-Term (E/P)
  • RBS-RFP (X/P)
  • pLacI (S/P)
  • TetR (X/P)

Gel Extraction


  • RBS-RFP
    • Concentration: 10.8 ng/μl
  • pLac
    • Concentration: 19.6 ng/μl
  • TetR
    • Concentration: 6.2 ng/μl

Ligation


  • Vector: pLac
  • Insert: TetR

Transformation


  • pLac-TetR
    • 0:1, 0 colonies seen
    • 2:1, 20 colonies seen, 1 picked
    • 4:1, 20 colonies seen, 1 picked

July 19


Triple Population and AND Gate Projects


Transformations


  • BBa_I721001: lead promoter
  • BBa_K091109: LuxS
  • BBa_C0078: LasI
  • BBa_C0062: LuxR
  • BBa_C0061: LuxI
  • BBa_E0432: RBS-YFP



July 20


Triple Population and AND Gate Projects


Colonies picked


  • LuxS: 20 colonies seen: 1 colony picked
  • LuxR: 5 colonies seen: 1 colony picked
  • LuxI: 5 colonies seen: 1 colony picked
  • RBS-YFP: no colonies seen
  • LasI: no colonies seen
  • lead promoter: no colonies seen


    • July 20


    Single and Double Population Projects


    Glycerol stocks & plasmid isolation


    • pLas-TetR: Concentrations: (2:1) 67.4 and (4:1) 68.0

    Digestion


    • (2:1) pLac-TetR

    Gel Extraction


    • pLac-TetR: Concentration: 20.5 ng/μl

    Ligation


    • Vector: pLac-TetR
    • Insert: RBS-YFP

    Transformation


    • pLac-TetR-RBS-YFP
      • (0:1) no colonies seen
  • (2:1) 1 colony picked

    • Single and Double Population Projects



    Triple Population and AND Gate Projects


    July 20


    Repeat Transformation


    • lead promoter
    • LasI
    • RBS-YFP

    Triple Population and AND Gate Projects


    July 21


    Glycerol Stock


    • LuxS
    • LuxR
    • LuxI

    Colonies picked


    • RBS-YFP: 1 colony picked
  • LasI: 1 colony picked
  • lead promoter: no colones seen

    • Digestion & Gel Extraction


    • LuxS (wrong enzymes used)
    • LuxR: Concentration: 11.1 ng/μl
    • LuxI: Concentration: 7.1 ng/μl

    Triple Population and AND Gate Projects


    July 22


    Plasmid Isolation


    • LuxS: Concentration: 75.2 ng/μl
    • LasI: Concentration: 90.1 ng/μl
    • RBS-YFP: Concentration: 76.4 ng/μl




    Triple Population and AND Gate Projects


    July 23


    DIgestion and gel extraction


    • LuxS (X/P) Concentration: not right size to be repeated
    • LasI (E/S) Concentration: 20.3 ng/μl
    • RBS-YFP (X/P) Concentration: 18.2 ng/μl
    • (wrong enzymes used in digestions - plasmid isolation repeated)

    Ligation


    • Vector: pLac
    • Insert: LuxR

    Transformation


    • BBa_ K117002: pLsrA
  • BBa_R0062: pLuxR
  • BBa_I721001: lead promoter

    • Colonies picked


    • pLac-LuxR: 15 colonies seen: 1 colony picked
    • pLsrA: 20 colonies seen: colony picked
    • pLuxR: 3 colonies seen: 1 colony picked



    July 23


    Glycerol stock and plasmid isolation


    • pLac-TetR-RBS-YFP: Concetration: 138.4

    Digestion


    • pLac-TetR-RBS-YFP (X/P)
    • pTetR-LacI-RBS-GFP (S/P)

    Ligation


    • Vector: pTetR-LacI-RBS-GFP
    • Insert: pLac-TetR-RBS-YFP
      • 2 colonies picked after plating and incubation

    Triple Population and AND Gate Projects


    July 24


    Plasmid Isolation


    • LuxS: Concentration: 85.9 ng/μl
    • LuxI: Concentration:85.7 ng/μl
    • LasI: Concentration: 224.1 ng/μl

    Digestion and gel extraction


    • LuxI (S/P): Concentration: 25.5 ng/μl
    • LasI (S/P): Concentration: 30.7 ng/μl

    Ligation


    • Vector 1: LuxI
    • Insert 1: RBS-YFP
    • Vector 2: LasI
    • Insert 2: RBS-RFP

    Triple Population and AND Gate Projects


    July 24


    Transformation


    • BBa_C0079: LasR
    • BBa_B0015: Term-Term
    • BBa_I14016: pLac
    • BBa_C0012: LacI

    Triple Population and AND Gate Projects


    July 25


    Colonies picked


    • LuxI-RBS-YFP: 100 colonies seen: 1 picked
    • LasI-RBS-RFP: 50 colonies seen: 1 picked

    Plasmid Isolation


    • pLac-LuxR: Concentration: 107.5
    • pLsrA: Concentration: 44.3
    • pLuxR: Concentration: 35.9

    Digestion and gel extraction


    • pLsrA (S/P): Concentration: 20.6 ng/μl
  • pLuxR (S/P): Concentration: 18.7 ng/μl

    • Transformation


    • BBa_R0040: pTetR
    • BBa_R0079: pLasR
    • BBa_K17036: RBS-CFP

    Single and Double Population Projects


    July 25


    Glycerol Stock and plasmid isolation


    • pTetR-LacI-RBS-GFP-pLac-TetR-RBS-YFP (full toggle switch) from both colines picked
      • Concentration 1: 194.8 ng/μl
      • Concentartion 2: 117.2 ng/μl

    Transformation


    • BBa_C0079: LasR
    • BBa_I14016: pLas
    • BBa_C0012: LacI

    Tetracycle Stock: 50 ng/чl


    IPTG stock: 50 ng/чl



    Triple Population and AND Gate Projects


    July 26


    Colonies picked


    • pTetR: 10 colonies seen: 1 colony picked
    • pLasR: 10 colonies seen: 1 colony picked
    • RBS-CFP: no colonies seen

    Glycerol stock and plasmid isolation


    • LuxI-RBS-YFP: Concentration: 70.8 ng/μl
    • LasI-RBS-RFP: Concentration: 4.0 (to be repeated) ng/μl

    Digestion and gel extraction


    • LuxI-RBS-YFP (X/P): Concentration: 2.8 (to be repeated)


    Single and Double Population Projects


    July 26


    Transformation


    • BBa_C0012: LacI
  • BBa_I732100: LacI (2)


    • Triple Population and AND Gate Projects


    July 27


    Glycerol stock and plasmid isolation


    • pTetR: Concentration: 42.4 ng/μl
    • pLasR: Concentration: 62.2 ng/μl
    • LuxI-RBS-YFP: Concentration: 120.9 ng/μl

    • LasI-RBS-RFP: Concentration: 85 ng/μl

    Digestion and gel extraction


    • pTetR (S/P): Concentration: 17.4 ng/μl
    • pLasR (S/P): Concentration: 23.8 ng/μl
    • LuxI-RBS-YFP (X/P): Concentration: 13.8 ng/μl

    • LasI-RBS-RFP (X/P): Concentration: 8.9 ng/μl

    Ligation


    • Vector 1: pLsrA
    • Insert 1:LuxI-RBS-YFP
  • Vector 2: pLuxR
  • Insert 2:LasI-RBS-RFP

    • Transformation:


    • pLsrA-LuxI-RBS-YFP
    • pLuxR-LasI-RBS-RFP

    • BBa_B0422: RBS-CFP



    Single and Double Population Projects


    July 27


    Colonies picked


    • LacI: 8 colonies seen: 1 colony picked
    • LacI (2): 2 colonies seen: 1 colony picked

    Plasmid isolation


    • LasR: Concentration: 95.8 ng/μl
    • Term-Term: Concentration: 120.1 ng/μl

    Toggle switch test


    • IPTG innoculation
  • TET innoculation
  • No innoculation


    • Triple Population and AND Gate Projects


    July 28


    Colonies picked


    • pLsrA-LuxI-RBS-YFP: 20 colonies seen: 1 colony picked
    • pLuxR-LasI-RBS-RFP: 2 colonies seen: 1 colony picked
    • RBS-CFP: 10 colonies seen: 1 colony picked


    Single and Double Population Projects


    July 28


    Glycerol stock and plasmid isolation


    • LacI: Concentration: 73.9 ng/μl
    • LacI (2): Concentration: 109.8 ng/μl

    Triple Population and AND Gate Projects


    July 29


    Glycerol Stocks


    • pLsrA-LuxI-RBS-YFP: Concentration: 201.6 ng/μl
    • pLuxR-LasI-RBS-RFP: Concentration: 127.1 ng/μl
    • RBS-CFP: Concentration: 84.0 ng/μl

    Triple Population and AND Gate Projects


    July 30


    Plasmid Isolation


    • pLsrA-LuxI-RBS-YFP: Concentration: 201.6 ng/μl
    • pLuxR-LasI-RBS-RFP: Concentration: 127.1 ng/μl
    • RBS-CFP: Concentration: 84.0 ng/μl

    Digestion and gel extraction


    • LuxS (X/P): Gel extraction failed
    • pLuxR-LasI-RBS-RFP (X/P): Gel extraction failed
    • RBS-CFP (X/P): Concentration: 10.4 ng/μl
    • pLac-LuxR (S/P): Concentration: 21.0 ng/μl

    Innoculation


    • pLuxR-LasI-RBS-RFP
    • LuxS

    Ligation and transformation:


    • Vector: pLasR
    • Insert: RBS-CFP
    • 2 colonies shown: 1 colony picked



    Single and Double Population Projects


    July 30


    Experiment 7


    • 150 ng/чl Tet
    • 200 ng/чl Tet
    • 300 ng/чl Tet
    • 1mM IPTG
    • No Induction

    Innoculation


    • RBS-GFP

    Triple Population and AND Gate Projects


    July 31


    Plasmid isolation


    • LuxS: Concentration: 78.1 ng/μl
    • pLuxR-LasI-RBS-RFP: Concentration: 48.7 ng/μl

    Digestion:


    • LuxS: (X/P); 516 bp

    • pLuxR+LasI-RBS-RFP: Digested at XbaI and PstI; 1421 bp

    Test Digestion:


    • LuxS: digested at EcoRI and PstI

    • pLuxR-lasI-RBS-RFP: digested at EcoRI and PstI

    Colonies:


    • pLuxR-lasI-RBS-RFP: 1 additional colony picked

    • LuxS: 1 additional colony picked


    Single and Double Population Projects


    July 31


    Plasmid Isolation:


    • GFP: 137.5 ng/μl



    August 1, 2012:


    Plasmid Isolation:


    • pLuxR-LasI-RBS-RFP: 61.2 ng/μl

    • pLasR-RBS-CFP: 49.2 ng/μl


    Digestion:


    • GFP: Digested XbaI and PstI

    • LacI: Digested XbaI and PstI

    • pLuxR-LasI-RBS-RFP: Digested XbaI and PstI

    • pLusR-RBS-CFP: Digested XbaI and PstI

    Gel Exraction:


    • GFP: 8.5 ng/μl

    • LacI: 12.7 ng/μl

    Colony:


    • pLasR-RBS-CFP: 1 additional colony picked

    Transformation:


    • pLuxR-lasI-RBS-RFP

    • BBa_J2309: pTetR-luxI



    Single and Double Population Projects


    August 1


    Ligation:


    • Vector: pTet; Insert: GFP

    • Vector: pTet; Insert: LacI

    • Vector: pLac; Insert: GFP

    Transformation:


    • pTet-GFP

    • pTet-LacI

    • pLac-GFP



    Triple Population and AND Gate Projects


    August 2, 2012:


    Colonies:


    • pLuxR-LasI-RBS-RFP: about 5 colonies; 1 picked

    • pTetR-LuxI: about 2 colonies; 1 picked

    Plasmid Isolation:


    • LuxS: Concentration – 40.5 ng/μl

    • pLasR-CFP: 95.8 ng/μl

    Digestion:


    • LasI: Digested at XbaI and PstI

    • LasR: Digested at XbaI and PstI

    • LuxS: Digested at XbaI and PstI

    • pLasR-RBS-CFP: Digested at XbaI and PstI

    Gel Extraction:


    • LasI: Concentration – 4.1 ng/μl

    • LasR: Concentration – 5.3 ng/μl

    • pLasR-RBS-CFP – 13.4 ng/μl

    Ligation:


    • Vector: pLac

    • Insert: LasI

    • Transformation 2:1 – 3 colonies; 1 picked



    Single and Double Population Projects


    August 2


    Colonies:


    • pTet-GFP: 7 colonies; 1 picked

    • pTet-LacI: 20 colonies; 1 picked

    • pLac-GFP: 20 colonies; 1 picked




    Single and Double Population Projects


    August 3, 2012:


    Glycerol Stocks/Miniprep:


    • pTet-GFP: Concentration-142.2 ng/μl

    • pTet-LacI: Concentration-67.2 ng/μl

    • pLac-GFP: Concentration-50.5 ng/μl

    Digestion:


    • pTet-GFP: Digested at SpeI and PstI

    • pTet-LacI: Digested at SpeI and PstI

    • pLac-GFP: Digested at XbaI and PstI

    • pLac-TetR: Digested at XbaI and PstI


    Triple Population and AND Gate Projects


    August 3


    Ligation:


    • Vector: pTetR

    • Insert: LasR

    • Transformation 2:1 – 1 colony; 1 picked

    Ligation:


    • Vector: pLasR

    • Insert: RBS-CFP

    • Transformation: 2:1

    Glycerol Stock:


    • pLuxR-LasI-RBS-RFP: Concentration – 45.1 ng/μl

    • pTetR-LuxI: Concentration – 105.1 ng/μl

    Digestion:


    • pLuxR-LasI-RBS-RFP: Digested at XbaI and PstI

    Ligation:


    • Vector: pLac-luxR
    • Insert: pLuxR-LasI-RBS-RFP

    Transformation


    • BBa_K091109 (2010) LuxS
    • BBa_K091109 (2011) LuxS

    Triple Population and AND Gate Projects


    August 4


    Colonies picked


    • BBa_K091109 (2010) LuxS: 70 colonies seen: 1 picked
    • BBa_K091109 (2011) LuxS: 100 colonies seen: 1 picked




    Triple Population and AND Gate Projects


    August 5


    Glycerol Stock


    • LuxS
    • LuxS
    • pTetR-RBS-CFP

    Transformation (repeated)


    • pLasR-RBS-CFP





    Triple Population and AND Gate Projects


    August 6


    Colonies picked


    • pLasR-RBS-CFP: 10 colonies seen: 1 colony picked

    Innoculation


    • pTetR-LuxI

    Plasmid isolation


    • LuxS: Concentration: 57.6
  • LuxS: Concentration: 76.4
  • pTetR-RBS-CFP: Concentration: 122.3

    • Digestion and gel extraction (improperly prepared for gel)


    • LuxS: Concentration: N/A
    • LuxS: Concentration: N/A
    • pTetR-RBS-CFP: Concentration: 21.9

    Innoculation


    • LuxS
    • LuxS
    • pTetR-RBS-CFP



    Single and Double Population Projects


    August 6, 2012:


    PCR:


    RFP-LVA created


    Experiments:


    Test mechanism shows TetR and LacI are medium strength repressors.


    Toggle Experiment fails to show significant changes in fluorescence protein after 2 or 6 hours.


    Triple Population and AND Gate Projects


    August 7



    Glycerol Stock


    • pLasR-RBS-CFP

    Plasmid isolation


    • LuxS: Concentration: 120.3
    • LuxS: Concentration:101.7
    • pTetR-LasR: Concentration: 65.3
    • pLasR-RBS-CFP: Concentration: 77.1
    • pLac-luxR-pLuxR-LasI-RBS-RFP: Concentration: 103.6

    Digestion and gel extraction


    • LuxS (X/P): Concentration: 1.5
    • LuxS (X/P): Concentration: 1.1
    • pTetR-LasR (S/P): Concentration: 20.4
  • pLasR-RBS-CFP (X/P): Concentration: 13.8

    • Ligation


    • Vector 1: pTetR-lasR
    • Insert 1: pLasR-RBS-CFP
    • Vector 2: pyear
    • Insert 2: LuxS

    Transformation


    • pSBIC3 chlora backbone
    • pLac-luxR-pLuxR-LasI-RBS-RFP



    Single and Double Population Projects


    August 7, 2012


    Glycerol Stock/Miniprep


    pTet-LacI


    pLac-GFP



    Digestion & Gel Extraction


    pTet-LacI: XbaI and PstI


    pLac-GFP: SpeI and PstI



    Ligation/Transformation


    Vector: pLac-GFP


    Insert: pTet-LacI




    Triple Population and AND Gate Projects


    August 8


    Colonies picked


    • chlora bb: 100 colonies seen: 1 picked
    • pLac-luxR-pLuxR-LasI-RBS-RFP: 300 colonies seen: 1 picked
    • pTetR-lasR-pLasR-RBS-CFP: 1 colony seen: 1 picked

    Repeat transformations from ligation


    • pyear-LuxS
    • pTetR-lasR-pLasR-RBS-CFP



    Single and Double Population Projects


    August 8, 2012



    Transformations:


    Negative control had ~20 colonies


    2:1 sample had ~ 40 colonies, we picked 2


    Positive had too many colonies to count



    PCR Reaction (Gradient)


    -55, 60, and 65 degrees

    -55 degrees found to be most ideal


    Gel Extraction (of PCR)


    Poor results, no DNA extracted


    Plate test for toggle switch:


    1 tube inoculated with IPTG 10 microliters of .2mM solution

    1 tube inoculated with ATC 10 microliters of 20ng/microliter


    Triple Population and AND Gate Projects


    August 9


    Glycerol stocks


    • chlora backbone
    • pLac-LuxR-pLuxR-LasI-RBS-RFP
  • pTetR-LasR-pLasR-RBS-CFP

    • Innoculation


    • pLac-Las-I
    • pTetR-LuxI

    Plasmid Isolation


    • chlora backbone

    • pLac-LuxR-pLuxR-LasI-RBS-RFP
    • pTetR-LasR-pLasR-RBS-CFP

    Digestion


    • Term-Term (S/P)

    • chlora backbone (E/P)

    • pLac-LuxR-pLuxR-LasI-RBS-RFP (E/P)
    • pTetR-LasR-pLasR-RBS-CFP (E/P)

    Gel Extraction


    • Term-Term: Concentration: 44.3

    • chlora backbone: Concentration: 332.9

    • pLac-LuxR-pLuxR-LasI-RBS-RFP: bad results
    • pTetR-LasR-pLasR-RBS-CFP: bad results

    Innoculation: pLac-LuxR-pLuxR-LasI-RBS-RFP




    Single and Double Population Projects


    August 9, 2012


    Glycerol stock/miniprep:


    pLac-GFP-ST-pTet-LacI



    PCR Reaction


    RFP-LVA



    Experiment: Repressor Test Mechanism (LacI)


    1 - Induced 10microliters of 10mM IPTG


    1 - No induction



    Digestion and Gel Extraction:


    RFP-LVA: XbaI and PstI


    Ampicillin Backbone: SpeI and PstI



    Triple Population and AND Gate Projects


    August 10


    Glycerol Stock


    • pLac-LuxR-pLuxR-LasI-RBS-RFP

    Plasmid isolation


    • pLac-LuxR-pLuxR-LasI-RBS-RFP: Concentration: 125.1

    Digestion


    • pLac-LuxR-pLuxR-LasI-RBS-RFP (E/P)

    Ligation


    • Vector1: Term-Term
    • Insert1: pLasR-RBS-CFP
    • Vector2: Term-Term
    • Insert2: pLuxR-LasI-RBS-RFP

    Transformation:


    • Term-Term-pLasR-RBS-CFP
    • Term-Term-pLuxR-LasI-RBS-RFP

    Single and Double Population Projects


    August 10, 2012


    Ligation/Transformation


    RFP-LVA into Ampicillin backbone



    Inoculations:


    Toggle


    pLas


    LacI


    LuxR


    pTet


    pLux


    LuxI



    August 11, 2012


    Plasmid isolation


    Toggle


    pLas


    LacI


    LuxR


    pTet


    pLux


    LuxI



    Digestion/Extraction


    LuxR: X and P


    pLux: S and P


    LuxI: S and P


    LasR: X and P


    Double Terminators (DT): X and P


    LasI: X and P




    Triple Population and AND Gate Projects


    August 11


    Colonies picked


    • Term-Term-pLasR-RBS-CFP: 200 colonies seen: 1 picked
    • Term-Term-pLuxR-LasI-RBS-RFP: 100 colonies seen: 1 picked

    Triple Population and AND Gate Projects


    August 12


    Glycerol Stock and Plasmid isolation


    • Term-Term-pLasR-RBS-CFP: Concentration: 219.1
    • Term-Term-pLuxR-LasI-RBS-RFP: Concentration: 107.5

    Transformation repeat


    • pyear-LuxS

    Digestion


    • Term-Term-pLasR-RBS-CFP (X/P)
    • Term-Term-pLuxR-LasI-RBS-RFP (X/P)

    Gel Extraction


    • Term-Term-pLasR-RBS-CFP: Concentration: 10.2
    • Term-Term-pLuxR-LasI-RBS-RFP: Concentration: 15.0

    Ligation


    • Vector1: pTetR-LasR
    • Insert1: Term-Term-pLasR-RBS-CFP
  • Vector2: pLac-LuxR
  • Insert 2: Term-Term-pLuxR-LasI-RBS-RFP

    • Triple Population and AND Gate Projects


    August 13


    Innoculation


    • pLac-LasI
    • pTetR-LuxI

    Single and Double Population Projects


    August 13, 2012


    Miniprep/Glycerol Stock


    RFP-LVA



    Digestions/Extraction


    pLas: S and P


    LacI: X and P


    RFP-LVA: E and P



    Inoculations


    Chlorabackbone


    RFP-LVA


    DT



    Triple Population and AND Gate Projects


    August 14


    Glycerol Stock and Plasmid isolation


    • pTetR-LasR-Term-Term-pLasR-RBS-CFP: Concentration: 95.4
    • pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP: Concentration: 108.3

    Digestion


    • pTetR-LasR-Term-Term-pLasR-RBS-CFP: bad results
    • pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP: bad results

    Parts Recieved


    • BBa_K091159
    • BBa_K091002
    • BBa_I13453
    • BBa_B0034

    Streaking


    • BBa_K091159 pBad-LsrA

    Ligation repeat


    • Vector1: pTetR-LasR
    • Insert1: Term-Term-pLasR-RBS-CFP
    • Vector2: pLac-LuxR
    • Insert 2: Term-Term-pLuxR-LasI-RBS-RFP

    Single and Double Population Projects


    August 14, 2012


    Miniprep/Glycerol Stock


    Chlorabackbone


    DT



    Digestion/Extraction:


    DT: X and P


    Chlorabackbone: E and P


    Toggle: E and P



    Ligation/Transformation:


    DT into LuxR


    TetR into pLux


    LasI into pTet


    LasR into DT


    pLas into LacI


    pLac into LuxI


    Triple Population and AND Gate Projects


    August 15


    Innoculation


    • pTetR-LuxI
    • pLac-LasI

    • pTetR-LasR-Term-Term-pLasR-RBS-CFP
    • pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP

    Colonies picked


    • pBad-LsrA: 30 colonies seen: 1 picked

    Testing


    • Pop2 (w/o TT) / pTetR-LuxI (48 hours of incubation)
      • 100/0 no results
      • 100/100 no results
      • 100/200 slight RFP detected no increase of RFP from 100/0
    • Pop3 (w/o TT) / pLac-LasI (48 hours of incubation)
      • 100/0 no results
      • 100/100 no results
      • 100/200 slight RFP detected no increase of CFP from 100/0

    Streaking


    BBa_K091109


    • SP3011>1D LuxS
    • SP4011>1D LuxS

    Single and Double Population Projects


    August 15, 2012


    Transformation


    LuxR-DT: 15 colonies, picked 1 colony


    pLux-TetR: 35 colonies, picked 1


    pTet-LasI: Negative control growth; unsuccessful


    LasR-DT: 25 colonies, picked 1


    pLas-LacI: 0 growth


    pLac-LuxI: 15 colonies, picked 1



    Ligation


    LasI into pTet



    Transformation


    pTet-LasI


    pLas-LacI



    Inoculations


    Chlorabackbone



    Triple Population and AND Gate Projects


    August 16


    Colonies


    • pTetR-LuxI:
    • pLac-LasI

    • pTetR-LasR-Term-Term-pLasR-RBS-CFP: 2 colonies seen: 1 colony picked
    • pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP: 0 colonies seen
    • LuxS: 200 colonies seen: 1 colony picked
    • LuxS: 100 colonies seen: 1 colony picked

    Glycerol Stock and plasmid isolation


    • pBad-LsrK: Concentration: 64.2
    • pTetR-LasR-Term-Term-pLasR-RBS-CFP: Concentration:
    • pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP: Concentration:

    Digestion


    • pTetR-LasR-Term-Term-pLasR-RBS-CFP (E/P)
    • pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP (E/P)
    • pBad-LsrA (S/P) 1749
    • pLsrA-LuxI-RBS-YFP (X/P)

    Testing


    • Pop2 (w/ TT) / pTetR-LuxI (6 hours of incubation)
      • 100/0
      • 100/200
      • 100/400
      • 100/600
      • Results: After 6 and 24 hours: no considerable difference
    • Pop3 (w/ TT) / pLac-LasI (6 hours of incubation)
      • 100/0
      • 100/200
      • 100/400
      • 100/600
      • Results: After 6 and 24 hours: no considerable difference
    • Pop3 (w/ TT) / Pop2 (w/ TT) / pTetR-LuxI (6 hours of incubation)
      • 100/0
      • 100/200
  • 100/400
  • 100/600
  • Results: After 6 and 24 hours: no considerable difference

    • Digestion


    • population 3 (oscillator)
    • population 2 (oscillator)
    • pBad->lsrK
    • pLsrA-LuxI-RBS-YFP
    • Term-Term (test)

    Gel Extraction


    • pBad-lsrK: Concentration: 53.8
    • pLsrA-LuxI-RBS-YFP: Concentration: 20.0

    Innoculation


    • Term-term

    Single and Double Population Projects


    August 16, 2012


    Miniprep/Glycerol Stock


    LuxR-DT


    pLux-TetR


    LasR-DT


    pLac-LacI


    Chlorabackbone


    Transformation Results


    Negative controls had too much growth; results thrown out



    Digestion/Extraction


    LuxR-DT: S and P


    pLux-TetR: X and P


    Chlorabackbone: E and P


    Ligations/Transformation


    LuxR-DT and pLux-TetR


    Chlorabackbone with Toggle Switch



    Triple Population and AND Gate Projects


    August 17


    Glycerol Stock


    • Population 3 w/ Term-Term
    • LuxS
    • LuxS

    Plasmid Isolation


    • Population 3 w/ Term-Term: Concentration: 126.7
    • LuxS: Concentration: 40.2
    • LuxS: Concentration: 52.4
    • Term-Term: Concentration: 100.8

    Digestion and Gel Extraction


    • Population 3 w/ Term-Term: Concentration: gel not used
    • LuxS: Concentration: gel not used
    • LuxS: Concentration: gel not used
  • Term-Term: Concentration: gel not used

    • Ligation


    • Vector 1: pLac
    • Insert 1: lasI-RBS-RFP
    • Vector 2: pLasR
    • Insert 2: LuxI-RBS-YFP
    • Vector 3: pbad-LsrK
    • Insert 3: pLsrA->YFP
    • Vector 4: pLac->LuxR
    • Insert 4: pLuxR-RFP
    • Vector 5: pTetR-LasR
    • Insert 5: pLasR->CFP

    Transformation


    • BBa_C0080: araC
    • BBa_J24674: pBad-TetR
    • BBa_I14015: pLas-TetO



    Single and Double Population Projects


    August 17, 2012


    Ligation


    pLas with LacI


    pTet with LasI



    Inoculations


    pLas


    LacI


    pTet


    pLac


    GFP


    RFP-LVA


    LasI



    Transformation


    Bba_K081014 Kit Plate 3 Well 12O Resistance AK; RFP



    Triple Population and AND Gate Projects


    August 18


    Innoculations


    • pTetR-LuxI
    • pLac-lasI
    • pop 2 w/TT (oscillator)
    • pop 3 w/TT (oscillator)

    Single and Double Population Projects


    August 18, 2012


    Colony Picking


    LuxR-DT-pLux-TetR: 30 colonies, 1 picked


    Toggle Switch in Chlorabackbone: 4 colonies, 1 picked


    pLas with LacI: negative control growth; results thrown out


    pTet with LasI: negative control growth; results thrown out


    RFP: 20 colonies, 1 picked


    Glycerol Stock


    LuxR-DT-pLux-TetR



    Minipreps


    LuxR-DT-pLux-TetR


    pLas


    LacI


    pTet


    pLac


    GFP


    LasI



    Triple Population and AND Gate Projects


    August 19


    Glycerol Stocks


    • pLac-lasI-RBS-RFP
    • pLasR-LuxI-RBS-YFP
    • pBad-LsrK-plsrA-LuxI-RBS-YFP
    • plac-LuxR-pLuxR-LasI-RBS-RFP
    • pTetR-lasR-pLasR-RBS-CFP
    • araC
    • pBad-TetR
    • pLas-TetO

    Oscillator Population Tests (population with induction plasmid)


    • pop2 0:1 - some fluorescence
    • pop2 75:1 - no visual difference
    • pop3 0:1 - some fluorescence
    • pop3 75:1 - no visual difference
    • pop2 and pop3 0:1 - little fluorescence
  • pop2 and pop3 75:1 - a visual increase of both RFP and CFP

    • Plasmid Isolation


    • pLac-lasI-RBS-RFP: Concentration: 115.8
    • pLasR-LuxI-RBS-YFP: Concentration: 93.1
    • pBad-LsrK-plsrA-LuxI-RBS-YFP: Concentration: 128.6
    • plac-LuxR-pLuxR-LasI-RBS-RFP: Concentration: 105.9
    • pTetR-lasR-pLasR-RBS-CFP: Concentration: 122.3
    • araC: Concentration: 114.0
    • pBad-TetR: Concentration: 315.1
    • pLas-TetO: Concentration: 102.3

    Digestion and gel extraction


    • ad-LsrK-plsrA-LuxI-RBS-YFP: Concentration:
    • plac-LuxR-pLuxR-LasI-RBS-RFP: Concentration:
    • pTetR-lasR-pLasR-RBS-CFP: Concentration:
    • araC: Concentration:
    • pBad-TetR: Concentration:
  • pLas-TetO: Concentration:
  • LasR-TT: Concentration: 10.0
  • LuxS: Concentration: 3.1
  • LuxS: Concentration: 5.5

    • Ligation


    • Vector 1: pyear
    • Insert 1: LuxS
    • Vector 2: pyear
    • Insert 2: LuxS
    • Vector 3: pTetR
    • Insert 3:lasR-TT
    • Vector 4: pLac-LuxR
    • Insert 4: TT
    • Vector 5: pLuxR
    • Insert 5: arac
    • Vector 6: pBad-TetR
  • Insert 6: TT
  • Vector 7: plasR-TetO
  • Insert 7: RBS-CFP

    • Single and Double Population Projects


    August 19, 2012


    Glycerol Stock/Plasmid Isolation


    RFP


    Toggle Switch in Chlorabackbone



    Inoculation


    RFP-LVA



    Digestion/Extraction


    pLac: S and P


    LuxR-DT-pLux-TetR: X and P


    pTet: S and P


    LasI: X and P


    LasR-DT: X and P


    LacI: S and P


    RFP: X and P



    Triple Population and AND Gate Projects


    August 20


    Colonies picked


    • pyear-LuxR: 1 picked
    • pyear-LuxR: 1 picked
    • pTet-lasR-TT: 1 picked
    • pLac-LuxR-TT: 1 picked
    • pBad-TetR-TT: 1 picked
    • pLuxR-arac: 1 picked
    • pLasR-TetO-RBS-CFP: 1 picked after being repeated

    Innoculation


    • pTetR-LuxI
    • pLac-lasI
    • pop3 w/TT
  • pop2 w/TT
  • pop3 w/o TT
  • pop2 w/o TT

    • Single and Double Population Projects


    August 20, 2012


    Ligations/Transformations


    LuxR-DT-pLux-TetR into pLac


    LasI into pTet


    LasR-DT into pTet


    RFP into LacI






    Triple Population and AND Gate Projects


    August 21


    Glycerol Stock and plasmid isolation


    • pyear-LuxR: Concentration: 40.0
    • pyear-LuxR: Concentration: 75.0
    • pTet-lasR-T: Concentration: 127.6
  • pLac-LuxR-TT: Concentration: 123.5
  • pBad-TetR-TT: Concentration: 141.0
  • pLuxR-arac: Concentration: 84.0

    • Digestion and gel extraction


    • pLasR-LuxI-YFP: gel not used
    • pTetR-lasR-TT: gel not used
    • pLuxR-araC: gel not used
    • pyear-LuxS: gel not used
    • pyear-LuxS: gel not used
    • pop1 (test)
    • pop2 (test)
    • pop3 (test)

    Innoculation


    • pyear-LuxS

    Ligation


    • Vector 1: pTet-LasR-TT
  • Insert 1: pLasR-LuxI-RBS-YFP
  • Vector 2: pLuxR-araC
  • Insert 2: lasR-TT

    • Single and Double Population Projects


    August 21, 2012


    Colony Pickings


    pLac-LuxR-DT-pLux-TetR: 24 colonies, 1 picked


    pTet-LasI: 30 colonies, 3 picked


    pTet-LasR-DT: 100 colonies, 2 picked


    LacI-RFP: 24 colonies, 2 picked



    Triple Population and AND Gate Projects


    August 22


    Digestion (test)


    • population 2 (oscillator)
      • undigested
      • EcoRI
  • XbaI
  • Spe1
  • Pst1
  • E/P
  • X/S

    • Innoculation


    • population 1 (oscillator)

    Colonies picked


    • pTetR-lasR-TT-pLasR-LuxI-RBS-YFP
    • pLuxR-araC-LasR-TT
    • pLasR-TetO-RBS-CFP (2 colonies picked)

    Transformation


    • pLac-LuxR

    Innoculation


    • pLuxR-LasI-RBS-RFP

    Population Testing Prep (pop2, pop3, pop2+pop3)


    • 2 ml of medium added to each test tube
    • 100 microliters of each population added as necessary
    • .75mL of inducing population added as necessary
      • pop2 w/ w/o inducer
      • pop3 w/ w/o inducer
      • pop2+pop3 w/ w/o inducer

    Single and Double Population Projects


    August 22, 2012


    Glycerol Stock/Miniprep


    pLac-LuxR-DT-pLux-TetR


    pTet-LasI


    pTet-LasR-DT


    LacI-RFP



    Digestion/Extraction:


    pLac-LuxR-DT-pLux-TetR: S and P


    GFP: X and P


    LacI-RFP: S and P


    pLac-LuxI: X and P


    Test Digestions


    pTet-LasI: E and P

    pTet-LasR-DT: E and P



    Ligations/Transformations


    GFP into pLac-LuxR-DT-pLux-TetR


    pLac-LuxI into LacI-RFP



    Inoculations


    RFP-LVA


    pTet-GFP


    RBS-YFP



    August 22


    Digestion (test)


    • population 2 (oscillator)
    • undigested
    • EcoRI
    • XbaI
    • Spe1
    • Pst1
    • E/P
    • X/S

    Innoculation


    • population 1 (oscillator)

    Colonies picked


    • pTetR-lasR-TT-pLasR-LuxI-RBS-YFP
    • pLuxR-araC-LasR-TT
    • pLasR-TetO-RBS-CFP (2 colonies picked)

    Transformation


    • pLac-LuxR

    Innoculation


    • pLuxR-LasI-RBS-RFP

    Population Testing Prep (pop2, pop3, pop2+pop3)


    • 2 ml of medium added to each test tube
    • 100 microliters of each population added as necessary
    • .75mL of inducing population added as necessary
      • pop2 w/ w/o inducer
      • pop3 w/ w/o inducer
      • pop2+pop3 w/ w/o inducer


    Single and Double Population Projects


    August 23, 2012



    Colony Picking


    LacI-RFP-ST-pLac-LuxI; 40 colonies, 1 picked


    LacI-RFP-ST-pLac-LuxI; 30 colonies, 1 picked




    Triple Population and AND Gate Projects


    August 23


    Glycerol Stock


    • pLasR-TetO-RBS-CFP (X2)
    • pTetR-lasR-TT-pLasR-LuxI-RBS-YFP
    • pLuxR-araC-LasR-TT

    Plasmid Isolation


    • pLasR-TetO-RBS-CFP: Concentration: 113.7
    • pLasR-TetO-RBS-CFP: Concentration: 112.0

    • pTetR-lasR-TT-pLasR-LuxI-RBS-YFP: Concentration: 167.0
    • pLuxR-araC-LasR-TT: Concentration: 104.2
    • pLac-lasI-RBS-RFP: Concentration: 37.0

    Colonies picked


    • pLac-LuxR

    Population Testing from 8/22 prep after 24 hours


    • 500 microliters from test tube of populations put in microcentrifuge tubes

      • pop2 w/: slight increase of RFP
      • pop2 w/o: some RFP
      • pop3 w/: no significant increase of CFP
  • pop3 w/o: some CFP
  • pop2+pop3 w/: no significant increase of RFP/CFP
  • pop2+pop3 w/o: some RFP/CFP
  • Results:
    • test tubes were checked after 24 hours
      • pop2 showed a slight increase in RFP production when it was induced in comparison to its uninduced control
      • both pop3 and pop2+pop3 showed no significant differences between its positive and negative controls
    • after 48 hours
      • pop2 showed increase in RFP production
      • pop3 showed no significant increase in CFP production
      • pop2+pop3 showed a sig increase in RFP production but no CFP when it was induced compared to its negative control

    Digestion


    • plac-LuxR(S/P): Concentration: 19.7
  • pTetR-LasR (E/P): Concentration: gel not used
  • pLasR-RBS-CFP (X/P): Concentration: gel not used
  • pLac-LuxR-TT (S/P): Concentration: 24.0
  • pBad-TetR-TT (S/P): Concentration: 15.4
  • pTetR-LasR-TT-pLasR-LuxI-RBS-YFP (E/P): Concentration: 12.2
  • pLuxR-araC-lasR-TT (X/P): Concentration: gel not used
  • pLasR-TetO-RBS-CFP (X/P): Concentration: 5.2
  • pLasR-TetO-RBS-CFP (X/P): Concentration: 5.2
  • pLuxR-LasI-RBS-RFP (X/P): Concentration: gel not used

    • Innoculation


    • pyear-luxS
    • pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
    • pTetR-LuxI
    • pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
    • pLac-lasI
    • pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
  • pLuxR-lasI-RBS-RFP

    • Ligation


    • Vector: pBad-TetR-TT
    • Insert: plasR-TetO-RBS-CFP

    Single and Double Population Projects


    August 24, 2012



    Plasmid Isolation


    LacI-RFP-ST-pLac-LuxI



    Digestion/Extraction


    pTet-LasR-DT


    pTet-LasI


    LacI-RFP-ST-pLac-LuxI


    pLas



    Ligation


    pTet-LasI into pLac-LuxR-DT-pLux-TetR-GFP-ST



    Triple Population and AND Gate Projects


    August 24


    Plasmid Isolation


    • pluxR-LasI-RBS-RFP: Concentration: 42.6

    Digestion


    • pLuxR-LasI-RBS-RFP (X/P)
    • pLuxR-araC-lasR-TT (X/P)

    Innoculation


    • pLuxR-LasI-RBS-RFP
    • pLuxR-araC-LasR-TT
    • pLsrA-LuxI-YFP



    Single and Double Population Projects


    August 25, 2012


    Glycerol Stock/Plasmid Isolation


    LacI-RFP-ST-pLac-LuxI



    Digestion/Extraction


    LacI-RFP-ST-pLac-LuxI



    Ligation


    LacI-RFP-ST-pLac-LuxI into pLas



    Triple Population and AND Gate Projects


    August 25


    Glycerol Stock


    • plac-LuxR
    • pBad-TetR-TT-plasR-TetO-RBS-CFP
    • pBad-TetR-TT-plasR-TetO-RBS-CFP
    • pLuxR-araC-lasR-TT

    Plasmid Isolation


    • pLuxR-LasI-RBS-RFP: Concentration: 89.1
    • pLuxR-araC-LasR-TT: Concentration: 152.9
    • pBad-TetR-TT-plasR-TetO-RBS-CFP: Concentration: 200.0
    • pBad-TetR-TT-plasR-TetO-RBS-CFP: Concentration: 156.0

    Digestion and Gel Extraction


    • pLuxR-LasI-RBS-RFP: (X/P): Concentration: 9.5
    • pLuxR-araC-LasR-TT: (X/P): gel not used
    • pBad-TetR-TT-plasR-TetO-RBS-CFP: (X/P): gel not used
    • pBad-TetR-TT-plasR-TetO-RBS-CFP: (X/P): gel not used

    Ligation


    • Vector1: pLac-LuxR-TT
    • Insert1: pLuxR-LasI-RBS-RFP
    • Vector2: pTetR-LasR
    • Insert2: pLasR-CFP
    • Vector3: pLac-LuxR-TT
    • Insert3: pLuxR-LasI-RBS-RFP

    Primer Dilution


    • I3.PsB1A2.R +661ul NFH2O -> 1:10 Dulition
    • I3.DT.F +634ul NFH2O -> 1:10 Dulition
    • I3.FT.M +700ul NFH2O -> 1:10 Dulition

    Colony PCR


    • pBad-TetR-TT-plasR-TetO-RBS-CFP
    • pLuxR-araC-LasR-TT
      • Primer 1: I3.24.SP1
      • Primer 2: I3.PSB1A2.R

    Innoculation


    • pyear-luxS
    • pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
    • pTetR-LuxI
    • pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
    • pLac-lasI
    • pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
    • pLuxR-lasI-RBS-RFP

    Triple Population and AND Gate Projects

    August 26, 2012


    Colony Picking


    pLas-LacI-RFP-ST-pLac-LuxI; 50 colonies, 1 picked



    Inoculation


    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP


    pLas-LacI-RFP-ST-pLac-LuxI



    Triple Population and AND Gate Projects


    August 26


    Colonies Picked


    • pLuxR-araC-LasR-TT: 1 colony picked

    Population testing


    • 2ml broth, 1ml induction, 100ul population, 30 hour incubation
      • pop1+2 (+/-): no fluorescence seen
      • pop2+3 (+/-): no fluorescence seen
      • pop1+2+3 (+/-): no fluorescence seen

    Single and Double Population Projects


    August 27, 2012


    Glycerol Stock/Plasmid Isolation


    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP


    pLas-LacI-RFP-ST-pLac-LuxI



    PCR


    Add second LacO site to pLac



    Digestion/Extraction


    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP


    pLas-LacI-RFP-ST-pLac-LuxI




    Triple Population and AND Gate Projects


    August 27


    Plasmid Isolation


    • pLuxR-araC-LaR-TT: Concentration: 164.3

    Digestion and Gel extraction


    • pLuxR-araC-LaR-TT (X/P): Concentration: 55.1

    Innoculation


    • pyear-luxS
    • pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
    • pTetR-LuxI
    • pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
    • pLac-lasI
    • pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)
    • pLuxR-lasI-RBS-RFP

    Ligation


    • Vector1: pBad-tetR-TT
    • Insert1: pLasR-TetO-RBS-CFP
      • 0:1 20 colonies seen
      • 2:1 50 colonies seen
    • Vector2:pLac-LuxR-TT
    • Insert2: pLuXR-araC-lasR-TT
    • 0:1 5 colonies seen
    • 2:1 25 colonies seen

    Single and Double Population Projects


    August 28, 2012


    Primer Dilutions



    Triple Population and AND Gate Projects


    August 28


    Colony PCR


    • pBad-tetR-TT-pLasR-TetO-RBS-CFP
    • pLac-LuxR-TT- pLuXR-araC-lasR-TT

    • Primer 1: I3.24.SP1
    • Primer 2: I3.PSB1A2.R

    Population Testing


    • 200ul medium, 200ul induction, 4ul population, 24 hour incubation

    • pop1+2 (-): no fluorescence seen
    • pop1+2 (+): some sign of RFP/YFP
  • pop2+3 (-): no fluorescence seen
  • pop2+3 (+): no fluorescence seen
  • pop1+2+3 (-): no fluorescence seen
  • pop1+2+3 (+): no fluorescence seen

    • Ligation


    • Vector:pLac-LuxR-TT
    • Insert: pLuXR-araC-lasR-TT
      • 0:1 0 colonies seen
      • 2:1 30 colonies seen
      • 4:1 15 colonies seen

    Digestion and Gel Extraction


    • pBad-TetR-TT (S/P): Concentration: 88.3

    Innoculation


    • pyear-luxS
    • pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
    • pTetR-LuxI
    • pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
  • pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)

    • Single and Double Population Projects


    August 29, 2012


    Ligation


    pLas-LacI-RFP-ST-pLac-LuxI into pTet-LasR-DT


    Triple Population and AND Gate Projects


    August 29


    Population Test Prep


    • 2ml medium, 400ul populations, 1500ul induction
      • pop1+2 (+/-)
      • pop2+3 (+/-)
      • pop1+2+3 (+/-)
    • 1ml medium, 200ul populations, 750ul induction
      • pop2+3 (+/-)

    Triple Population and AND Gate Projects


    August 30


    Ligation


    • Vector: pBad-TetR-TT
    • Insert: plasR-TetO-CFP
      • 0:1 100 Colonies seen
      • 1:1 200 Colonies seen
      • 4:1 200 Colonies seen

    Colony PCR


    • pLac-LuxR-TT-pLuXR-araC-lasR-TT (X2)

    • Primer 1: I3.24.SP1
    • Primer 2: I3.PSB1A2.R

    Colonies picked


    • pLac-LuxR-TT-pLuXR-araC-lasR-TT (X2)

    Single and Double Population Projects


    August 31, 2012


    Colony picking


    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI; 50 colonies, 1 picked



    PCR Reaction


    Add LacO binding site to pLac; 65.2 degrees found to be optimal temperature for elongation



    Transformation


    pLac-LacO


    Triple Population and AND Gate Projects


    August 31



    Glycerol Stock and Plasmid Isolation


    • pLac-LuxR-TT-pLuXR-araC-lasR-TT: Concentration: 115.2
    • pLac-LuxR-TT-pLuXR-araC-lasR-TT: Concentration: 77.7

    Digestion and Gel Extraction


    • pLac-LuxR-TT-pLuXR-araC-lasR-TT (S/P)
  • pLac-LuxR-TT-pLuXR-araC-lasR-TT (S/P)

    • Colony PCR


    • pBad-TetR-TT-pLasR-TetO-CFP (X2)

    • Primer 1: I3.24.SP1
    • Primer 2: I3.PSB1A2.R

    Population Test Results


    • 2ml medium, 400ul populations, 1500ul induction
      • pop1+2 (+/-)
      • pop2+3 (+/-)
      • pop1+2+3 (+/-)
    • 1ml medium, 200ul populations, 750ul induction
      • pop2+3 (+/-)

    Single and Double Population Projects


    September 1, 2012


    Glycerol Stock/Plasmid Isolation


    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI



    Colony Picking


    pLac-LacO



    Inoculations


    pLac-TetR


    pLac-TetR-RBS-RFP


    pTet-LacI


    pTet-RBS-GFP-T



    Triple Population and AND Gate Projects


    September 1


    Ligations


    • Vector1: pLac-LuxR-TT
    • Insert1: pLuxR-araC-LasR-TT
    • Vector2: pLasR
    • Insert2.1: LuxS
    • Insert2.2 araC
    • Vector3: TT
  • Insert3: pLsrA-LuxI-RBS-YFP
  • Vector4: pLuxR
  • Insert4: TetR

    • Transformation


    • BBa_C0012: LacI
    • BBa_K091002: LsrK

    Innoculation


    • pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)


    Single and Double Population Projects


    September 2, 2012


    Glycerol Stock/Plasmid Isolation


    pLac-LacO



    Single and Double Population Projects


    September 3, 2012


    Plasmid Isolation


    pLac-TetR


    pLac-TetR-RBS-RFP


    pTet-LacI


    pTet-RBS-GFP-T



    Digestion/Extraction


    pLac-TetR

    pLac-TetR-RBS-RFP


    pTet-LacI


    pTet-RBS-GFP-T


    Triple Population and AND Gate Projects


    September 3


    Glycerol Stock


    • pLac-LuxR-TT-pLuxR-araC-LasR-TT
    • pBad-TetR-TT-pLas-TetO-CFP

    Plasmid Isolation


    • pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
    • pLac-LuxR-TT-pLuxR-araC-lasR-TT
    • pBad-TetR-TT-pLas-TetO-CFP

    Digestion


    • pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator) (X/P)
    • pBad-TetR-TT-pLas-TetO-CFP (X/P)
    • pLac-LuxR-TT-pLuxR-araC-lasR-TT (S/P)

    Innoculation


    • pyear-luxS (Inducer pop1)
    • pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
  • pTetR-LuxI (Inducer pop2)
  • pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
  • pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)

    • Colony PCR for I3-98-2 and I4-1-2-


    Digestion


    • TT-pLuxR-LasI-RFP (X/P)
    • pLac-LuxR (S/P)

    Single and Double Population Projects


    September 4, 2012


    Inoculations


    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI (Atc 10 microliters)


    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI (IPTG 10 microliters)



    Triple Population and AND Gate Projects


    September 4


    Plasmid Isolation


    • pBad-TetR-TT-pLasR-TetO-CFP: Concentration: 190.4

    Digestion


    • pBad-TetR-TT-pLasR-TetO-CFP (X/P): Concentration: 20.5

    Colonies picked from I4-1-2




    Single and Double Population Projects


    September 5, 2012


    Inoculations


    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI

    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI


    1 with Atc


    1 with IPTG


    1 with No induction


    Triple Population and AND Gate Projects


    September 5


    Glycerol Stock and Plasmid Isolation


    • pLasR-LuxS
    • pLasR-araC
    • TT-pLsrA-LuxI-RBS-YFP
  • pLuxR-TetR
  • LacI
  • LsrK

    • Digestion and Gel Extraction


    • pLasR-LuxS
    • pLasR-araC
    • TT-pLsrA-LuxI-RBS-YFP
    • pLuxR-TetR
    • LacI
    • LsrK

    Innoculation


    • pLac
    • LasI-RFP

    Single and Double Population Projects


    September 6, 2012


    Sequencing


    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI


    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI


    pLac-LacO


    RFP-LVA



    Inoculations


    Chlorabackbone


    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI

    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI


    1 with Atc


    1 with IPTG


    1 with No induction



    Digestion/Extraction


    Chlorabackbone



    Triple Population and AND Gate Projects


    September 6


    Plasmid Isolation and Glycerol Stock


    • pLac
    • LasI-RFP
    • pLac-LuxR-TT-pLuxR-araC-LasR-TT

    Digestion and Gel Extraction


    • pLac
    • LasI-RFP
    • pLac-LuxR-TT-pLuxR-araC-LasR-TT

    Ligation


    • Vector1:pLac-LuxR-TT-pLuxR-araC-LasR-TT
    • Insert1: pBad->>CFP
    • Vector2: pLsrA
    • Insert2: LacI
    • Vector3: pLasR-LuxS
  • Insert3: RBS-CFP
  • Vector4: Term-Term
  • Insert4: pLuxR-TetR

    • Single and Double Population Projects


    September 7, 2012


    Inoculation


    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP


    pLac-RBS-GFP-T


    pTet-RBS-GFP-T-pLac-TetR

    pLac-RBS-GFP-T-pTet-LacI

    RFP-LVA

    pLac-LacO

    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI

    pTet-LasI

    pLac-LuxR-DT-pLux-TetR

    LuxR-DT-pLux-TetR

    LuxR-DT

    pLux-TetR

    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI

    pLas-LacI-RFP-ST-pLac-LuxI

    LacI-RFP-ST-pLac-LuxI

    LacI-RFP-ST

    pLac-LuxI

    pTet-LasR-DT

    LasR-DT

    pLac-LacO

    pTet-LacI

    RBS-GFP-ST


    Triple Population and AND Gate Projects


    September 7


    Test Digestions


    • pLac
    • LasI-RFP
  • pLac-LuxR-TT-pLuxR-araC-LasR-TT

    • Innoculation


    • Term-Term

    Single and Double Population Projects


    September 8, 2012


    Time-Lapse Experiment to observe toggle switch


    Plasmid Isolation


    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP

    pLac-RBS-GFP-T


    pTet-RBS-GFP-T-pLac-TetR

    pLac-RBS-GFP-T-pTet-LacI

    RFP-LVA

    pLac-LacO

    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI

    pTet-LasI

    pLac-LuxR-DT-pLux-TetR

    LuxR-DT-pLux-TetR

    LuxR-DT

    pLux-TetR

    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI

    pLas-LacI-RFP-ST-pLac-LuxI

    LacI-RFP-ST-pLac-LuxI

    LacI-RFP-ST

    pLac-LuxI

    pTet-LasR-DT

    LasR-DT

    pTet-LacI

    GFP


    Ligations


    pLac-TetR into Chlora


    pLac-RBS-GFP-T into Chlora


    pTet-RBS-GFP-T into Chlora


    pTet-LacI into Chlora



    ]

    Digestion/Extraction


    GFP


    pLac-LacO


    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP

    pLac-RBS-GFP-T


    pTet-RBS-GFP-T-pLac-TetR

    pLac-RBS-GFP-T-pTet-LacI

    RFP-LVA

    pLac-LacO

    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI

    pTet-LasI

    pLac-LuxR-DT-pLux-TetR

    LuxR-DT-pLux-TetR

    LuxR-DT

    pLux-TetR

    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI

    pLas-LacI-RFP-ST-pLac-LuxI

    LacI-RFP-ST-pLac-LuxI

    LacI-RFP-ST

    pLac-LuxI


    Triple Population and AND Gate Projects


    September 8


    Colonies Picked


    • 14-7-5
    • 14-7-7
    • 14-8-2

    Single and Double Population Projects


    September 9, 2012


    Colony Picking


    pLac-TetR, 1 colony picked


    pLac-RBS-GFP-T, 1 colony picked


    pTet-LacI, 1 colony picked



    Ligation


    pLac-LacO-GFP



    Triple Population and AND Gate Projects


    September 9


    Glycerol Stock and Plasmid Isolation


    • pLac-LuxR-TT-pLuxR-araC-LasR-TT-pBad-TetR-TT-pLasR-TetO-CFP
    • pLsrA-LacI
    • pLasR-LuxS-RBS-CFP

    Digestion and Gel Extraction


    • pLac-LuxR-TT-pLuxR-araC-LasR-TT-pBad-TetR-TT-pLasR-TetO-CFP
  • pLsrA-LacI
  • pLasR-LuxS-RBS-CFP

    • Ligations


    • Vector1: Term-Term
    • Insert1-1: pLsrA-LacI
    • Insert1-2: pLuxR-TetR
    • Insert1-3: pLsrA-LuxI-RBS-YFP
    • Insert1-4: pLasR-LuxS-RBS-CFP
    • Insert1-5: pLasR-araC
    • Vector2: pLac
    • Insert2: 13-12-8

    Single and Double Population Projects


    September 10, 2012


    Glycerol Stock/Miniprep


    pLac-TetR


    pLac-RBS-GFP-T


    pTet-LacI



    Gel Extraction


    pLac-LacO-GFP


    pTet-LacI



    Ligation


    pTet-LacI into pLac-LacO-RBS-GFP-ST


    pLac-RBS-GFP-T-pTet-LacI into chlora backbone



    Triple Population and AND Gate Projects


    September 10


    Colonies Picked


    • I4-11-2
    • I4-11-3
    • I4-11-4
    • I4-11-5
    • I4-11-6
    • I4-8-4
  • I4-8-5
  • I4-8-6

    • Plasmid Isolation


    • pLac
    • LuxR

    Single and Double Population Projects


    September 11, 2012


    PCR


    pLac-LacO


    pLac-LacO x2



    Single and Double Population Projects


    September 12, 2012


    Digestion/Gel Extraction


    pTet-LacI-pLac-LacO-RBS-GFP-ST



    Experiments


    Toggle with 20 micro liters IPTG


    Toggle with 40 micro liters ATc


    Strain 1 of dual population toggle with inducer strain


    Strain 2 of dual population toggle with inducer strain



    Ligation


    pLac-LacO-RBS-GFP-ST-pTet-LacI into chlora backbone



    Triple Population and AND Gate


    September 13


    Glycerol Stock and Plasmid Isolation


    • I4-8-4
    • I4-8-5
    • I4-8-6
    • I4-11-2
    • I4-11-3
    • I4-11-4
    • I4-11-5
  • I4-11-6
  • I4-13-1
  • I4-13-2

    • Innoculation


    • pyear-luxS
    • pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
    • pTetR-LuxI
    • pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
    • pLac-lasI
    • pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)

    Ligation


    • Vector: Const. Promoter
    • Insert1-1: I3-28-4
    • Insert1-2: I4-6-6
    • Insert1-3: I4-16-2
    • Vector2: TT-pLasR-LuxS-RBS-CFP
  • Insert2: TT-pLuxR-TetR
  • Vector3: pLac
  • Insert3: LuxR
  • Vector4: pLuxR
  • Insert4: LasI-RFP

    • Ligations for Chlora


    • I4-14-7
    • I4-14-8
    • I4-14-10

    Transformation


    • I4-18-1
    • I4-18-2
    • I4-18-3
    • I401804

    Innoculation


    • I3-28-2
  • I3-30-4
  • I3-11-1
  • I3-46-4

    • Single and Double Population Projects


    September 14, 2012


    Ligation


    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP into Chlora backbone

    pLac-RBS-GFP-T into Chlora backbone


    pTet-RBS-GFP-T-pLac-TetR into Chlora backbone

    pLac-RBS-GFP-T-pTet-LacI into Chlora backbone

    RFP-LVA into Chlora backbone

    pLac-LacO into Chlora backbone

    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI into Chlora backbone

    pTet-LasI into Chlora backbone

    pLac-LuxR-DT-pLux-TetR into Chlora backbone

    LuxR-DT-pLux-TetR into Chlora backbone

    LuxR-DT into Chlora backbone

    pLux-TetR into Chlora backbone

    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI into Chlora backbone

    pLas-LacI-RFP-ST-pLac-LuxI into Chlora backbone

    LacI-RFP-ST-pLac-LuxI into Chlora backbone

    LacI-RFP-ST into Chlora backbone

    pLac-LuxI into Chlora backbone

    pLac-LacO x2-RBS-GFP-ST-pTet-LacI


    Glycerol Stock/Plasmid Isolation


    pLac-LacO-GFP



    Digestion


    pLac-LacO-GFP


    pTet-LacI


    pLac-LacO x2-RBS-GFP-ST-pTet-LacI


    Inoculation


    pLac-GFP-pTet-LacI



    Triple Population and AND Gate


    September 15


    Plasmid Isolation


    • I4-16-4 CP-LasR
    • I4-16-5 CP-LsrK
    • I4-16-6 CP-LuxR
    • I4-17-4 TT-pLasR->>TetR
    • I4-17-6 pLac-LuxR
    • I4-17-8 pLuxR-LasI-RFP

    Colonies picked


    • I4-19-19
    • I4-22-8
    • I4-22-9
    • I4-21-2

    Digestion


    • I4-16-4
  • I4-16-5
  • I4-16-6
  • I4-17-4
  • I4-17-6
  • I4-17-8

    • Ligations


    • Vector1: pTetR-LasR
    • Insert1: TT-pLasR-LuxS-RFP-TT-pLuxR-TetR
    • Vector2: Term-Term
    • Insert2: pLac-LuxR
    • Vector3: pLuxR-LasII-RFP
    • Insert3: TT-pLsrA-LacI

    Innoculation


    • pyear-luxS
    • pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
    • pTetR-LuxI
  • pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
  • pLac-lasI
  • pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)



    • Triple Population and AND Gate


    September 18


    Glycerol Stock and Plasmid Isolation


    • pTetR-LasR-TT-pLasR-LuxS-CFP-TT-pLuxR-TetR
    • TT-pLac-LuxR
    • pLuxR-LasI-RFP-TT-pLsrA-LacI

    Digestion and Gel Extraction


    • I4-10-7
    • I4-25-14
    • I4-25-16
    • I4-25-18

    Ligation


    • Vector1: CP-LasR
    • Insert1: TT
    • Vector2: TT-pLsrA-LuxI-YFP
    • Insert 2: TT-pLasR-araC
  • Vector3: TT
  • Insert3.1: pTetR->>TetR
  • Insert3.2: pLuxR->>LacI
  • Vector4: CP-LsrK
  • Insert4: TT-pLac-LuxR

    • Innoculation


    • Pop3 AND Gate
    • Pop2 AND Gate
    • Pop1 AND Gate

    Testing AND Gate


    • Pop2 and 3
    • Pop1 and 3
    • Pop 1 2 3

    Triple Population and AND Gate


    September 19


    Plate Experiments for AND Gate







    Triple Population and AND Gate


    September 20


    Glycerol Stock and Plasmid Isolation


    • Cp-LasR-TT
    • TT-pLsrA->>araC
    • TT-pTetR->>TetR
    • CP-LsrK->>LuxR
    • TT-pLuxR->>LacI

    Digestions (repeated 3X)


    • Cp-LasR-TT
    • TT-pLsrA->>araC
    • TT-pTetR->>TetR
    • CP-LsrK->>LuxR
    • TT-pLuxR->>LacI

    Triple Population and AND Gate


    September 22


    Ligations (Repeated)


    • Vector1: CP-LasR
    • Insert1: TT
    • Vector2: TT-pLsrA-LuxI-YFP
    • Insert 2: TT-pLasR-araC
    • Vector3: TT
    • Insert3.1: pTetR->>TetR
    • Insert3.2: pLuxR->>LacI
    • Vector4: CP-LsrK
    • Insert4: TT-pLac-LuxR

    Single and Double Population Projects


    September 22, 2012


    Single toggle experiment - No induction at start, check every 6 hours, induce with IPTG after first 6 hours, and induce with ATc in new tube after 12 Hours


    Triple Population and AND Gate


    September 22


    Ligations (Repeated)


    • Vector1: CP-LasR
    • Insert1: TT
    • Vector2: TT-pLsrA-LuxI-YFP
    • Insert 2: TT-pLasR-araC
    • Vector3: TT
    • Insert3.1: pTetR->>TetR
    • Insert3.2: pLuxR->>LacI
    • Vector4: CP-LsrK
    • Insert4: TT-pLac-LuxR

    Triple Population and AND Gate


    September 23


    Digestions


    • Cp-LasR-TT
    • TT-pLsrA->>araC
    • TT-pTetR->>TetR
    • CP-LsrK->>LuxR
    • TT-pLuxR->>LacI

    Innoculations for AND Gate


    • Pop1
    • Pop2
    • Pop3

    Testing AND Gate


    • Pop 1 3
    • Pop 2 3
    • Pop 1 2 3

    Innoculation


    • pyear-luxS
    • pBad-LsrK-pLsaA-LuxI-RBS-YFP (pop1 oscillator)
    • pTetR-LuxI
    • pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
    • pLac-lasI
    • pTetR-lasR-TT-pLasR-RBS-CFP (pop3 oscillator)


    Triple Population and AND Gate


    September 23


    Plasmid Isolation


    • TT-pLsrA-LuxI->>araC
    • TT-pTetR-TetR
    • CP-LsrK->>LuxR
    • TT-pLuxR-LacI

    Digestions and Gel Extraction


    • TT-pLsrA-LuxI->>araC
    • TT-pTetR-TetR
    • CP-LsrK->>LuxR
    • TT-pLuxR-LacI

    Retrieved from "http://2012.igem.org/Team:UT_Dallas/notebook_data"