Team:UT Dallas/notebook data

From 2012.igem.org

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<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>
 +
<p>
<p>
-
Transformations
+
    Single and Double Population Projects
</p>
</p>
 +
<p>
<p>
-
&#9679; BBa_I721001: lead promoter
+
    Transformation
</p>
</p>
 +
 +
<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>
 +
<p>
<p>
-
&#9679; BBa_K091109: LuxS
+
    Single and Double Population Projects
</p>
</p>
 +
<p>
<p>
-
&#9679; BBa_C0078: LasI
+
    June 7
</p>
</p>
 +
<p>
<p>
-
&#9679; BBa_C0062: LuxR
+
    Colonies picked and glycerol stocked made
</p>
</p>
 +
 +
<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>
 +
 +
</div>
 +
<div id="page3" class='notebook_page'>
<p>
<p>
-
&#9679; BBa_C0061: LuxI
+
    Single and Double Population Projects
</p>
</p>
 +
<p>
<p>
-
&#9679; BBa_E0432: RBS-YFP
+
    June 8
</p>
</p>
 +
<p>
<p>
-
<strong>July 20</strong>
+
    Plasmid isolation
</p>
</p>
 +
 +
<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>
 +
 +
</div>
 +
<div id="page4" class='notebook_page'>
 +
<p>
<p>
-
Colonies picked
+
    Single and Double Population Projects
</p>
</p>
 +
<p>
<p>
-
&#9679; LuxS: 20 colonies seen: 1 colony picked
+
    June 11
</p>
</p>
 +
<p>
<p>
-
&#9679; LuxR: 5 colonies seen: 1 colony picked
+
    Digestion
</p>
</p>
 +
 +
<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>
 +
<p>
<p>
-
&#9679; LuxI: 5 colonies seen: 1 colony picked
+
    Single and Double Population Projects
</p>
</p>
 +
<p>
<p>
-
&#9679; RBS-YFP: no colonies seen
+
    June 12
</p>
</p>
 +
<p>
<p>
-
&#9679; LasI: no colonies seen
+
    Digestion
</p>
</p>
 +
 +
<ul>
 +
    <li>
 +
        RBS-GFP (X/P)
 +
    </li>
 +
</ul>
<p>
<p>
-
&#9679; lead promoter: no colonies seen
+
    Ligation
</p>
</p>
 +
 +
<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>
 +
 +
</div>
 +
<div id="page5" class='notebook_page'>
<p>
<p>
-
<strong>July 20</strong>
+
    June 13
</p>
</p>
 +
<p>
<p>
-
Repeat Transformation
+
    Incubate constitutive promoter-RBS-GFP in LB carb broth overnight @ 37*C and 220 rpm
</p>
</p>
 +
 +
<ul>
 +
    <li>
 +
        2:1--- 2 colonies picked
 +
    </li>
 +
</ul>
 +
<p>
<p>
-
&#9679; lead promoter
+
    Single and Double Population Projects
</p>
</p>
 +
<p>
<p>
-
&#9679; LasI
+
    June 14
</p>
</p>
 +
<p>
<p>
-
&#9679; RBS-YFP
+
    Glycerol stock and miniprep
</p>
</p>
 +
 +
<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>
 +
 +
<p>
<p>
-
Glycerol Stock
+
    Single and Double Population Projects
</p>
</p>
 +
<p>
<p>
-
&#9679; LuxS
+
    June 15
</p>
</p>
 +
<p>
<p>
-
&#9679; LuxR
+
    Plasmid isolation
</p>
</p>
 +
 +
<ul>
 +
    <li>
 +
        prom-RBS-GFP: Concentration: 127 ng/µl
 +
    </li>
 +
</ul>
 +
 +
</div>
 +
<div id="page6" class='notebook_page'>
 +
<p>
<p>
-
&#9679; LuxI
+
    Single and Double Population Projects
</p>
</p>
 +
<p>
<p>
-
Colonies picked
+
    July 16
</p>
</p>
 +
<p>
<p>
-
&#9679; RBS-YFP: 1 colony picked
+
    Transformations
</p>
</p>
 +
 +
<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>
 +
<p>
<p>
-
&#9679; LasI: 1 colony picked
+
    Single and Double Population Projects
</p>
</p>
 +
<p>
<p>
-
&#9679; lead promoter: no colones seen
+
    July 17
</p>
</p>
 +
 +
<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'>
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    July 18
 +
</p>
 +
 +
<p>
 +
    Glycerol stocks made
 +
</p>
 +
 +
<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>
 +
    Digestion
 +
</p>
 +
 +
<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>
 +
 +
</div>
 +
<div id="page8" class='notebook_page'>
 +
 +
<p>
 +
    Gel Extraction
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Vector: pLac
 +
    </li>
 +
    <li>
 +
        Insert: TetR
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Transformation
 +
</p>
 +
 +
<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>
 +
 +
</div>
 +
<div id="page9" class='notebook_page'>
 +
 +
<p>
 +
    July 19
 +
</p>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    Transformations
 +
</p>
 +
 +
<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>
 +
 +
 +
 +
<p>
 +
    July 20
 +
</p>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<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>
 +
 +
 +
<p>
 +
    July 20
 +
</p>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    Glycerol stocks &amp; plasmid isolation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLas-TetR: Concentrations: (2:1) 67.4 and (4:1) 68.0
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Digestion
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        (2:1) pLac-TetR
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Gel Extraction
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-TetR: Concentration: 20.5 ng/µl
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Vector: pLac-TetR
 +
    </li>
 +
    <li>
 +
        Insert: RBS-YFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Transformation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-TetR-RBS-YFP
 +
    </li>
 +
    <ul>
 +
        <li>
 +
            (0:1) no colonies seen
 +
        </li>
 +
 +
</div>
 +
<div id="page11" class='notebook_page'>
 +
 +
        <li>
 +
            (2:1) 1 colony picked
 +
        </li>
 +
    </ul>
 +
</ul>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    July 20
 +
</p>
 +
 +
<p>
 +
    Repeat Transformation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        lead promoter
 +
    </li>
 +
    <li>
 +
        LasI
 +
    </li>
 +
    <li>
 +
        RBS-YFP
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    July 21
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
    <li>
 +
        LuxR
 +
    </li>
 +
    <li>
 +
        LuxI
 +
    </li>
 +
</ul>
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<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>
 +
    Digestion &amp; Gel Extraction
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LuxS (wrong enzymes used)
 +
    </li>
 +
    <li>
 +
        LuxR: Concentration: 11.1 ng/µl
 +
    </li>
 +
    <li>
 +
        LuxI: Concentration: 7.1 ng/µl
 +
    </li>
 +
</ul>
 +
 +
</div>
 +
<div id="page13" class='notebook_page'>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
 +
 +
    July 22
 +
</p>
 +
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<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>
 +
 +
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    July 23
 +
</p>
 +
 +
<p>
 +
    DIgestion and gel extraction
 +
</p>
 +
 +
<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>
 +
    Ligation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Vector: pLac
 +
    </li>
 +
    <li>
 +
        Insert: LuxR
 +
    </li>
 +
</ul>
 +
<p>
 +
    Transformation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        BBa_ K117002: pLsrA
 +
    </li>
 +
 +
</div>
 +
<div id="page14" class='notebook_page'>
 +
 +
    <li>
 +
        BBa_R0062: pLuxR
 +
    </li>
 +
    <li>
 +
        BBa_I721001: lead promoter
 +
    </li>
 +
</ul>
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<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>
 +
 +
 +
 +
<p>
 +
    July 23
 +
</p>
 +
 +
<p>
 +
    Glycerol stock and plasmid isolation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-TetR-RBS-YFP: Concetration: 138.4
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-TetR-RBS-YFP (X/P)
 +
    </li>
 +
    <li>
 +
        pTetR-LacI-RBS-GFP (S/P)
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<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>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    July 24
 +
</p>
 +
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<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>
 +
    Digestion and gel extraction
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LuxI (S/P): Concentration: 25.5 ng/µl
 +
    </li>
 +
    <li>
 +
        LasI (S/P): Concentration: 30.7 ng/µl
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Vector 1: LuxI
 +
    </li>
 +
    <li>
 +
        Insert 1: RBS-YFP
 +
    </li>
 +
    <li>
 +
        Vector 2: LasI
 +
    </li>
 +
    <li>
 +
        Insert 2: RBS-RFP
 +
    </li>
 +
</ul>
 +
 +
 +
</div>
 +
<div id="page16" class='notebook_page'>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    July 24
 +
</p>
 +
 +
<p>
 +
 +
    Transformation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        BBa_C0079: LasR
 +
    </li>
 +
    <li>
 +
        BBa_B0015: Term-Term
 +
    </li>
 +
    <li>
 +
        BBa_I14016: pLac
 +
    </li>
 +
    <li>
 +
        BBa_C0012: LacI
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    July 25
 +
</p>
 +
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<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>
 +
 +
<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>
 +
 +
<ul>
 +
    <li>
 +
        pLsrA (S/P): Concentration: 20.6 ng/µl
 +
    </li>
 +
 +
</div>
 +
<div id="page17" class='notebook_page'>
 +
 +
    <li>
 +
        pLuxR (S/P): Concentration: 18.7 ng/µl
 +
    </li>
 +
</ul>
 +
<p>
 +
    Transformation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        BBa_R0040: pTetR
 +
    </li>
 +
    <li>
 +
        BBa_R0079: pLasR
 +
    </li>
 +
    <li>
 +
        BBa_K17036: RBS-CFP
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    July 25
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock and plasmid isolation
 +
</p>
 +
 +
<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>
 +
    Transformation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        BBa_C0079: LasR
 +
    </li>
 +
    <li>
 +
        BBa_I14016: pLas
 +
    </li>
 +
    <li>
 +
        BBa_C0012: LacI
 +
    </li>
 +
</ul>
 +
<p>
 +
    Tetracycle Stock: 50 ng/?l
 +
</p>
 +
 +
 +
</div>
 +
<div id="page18" class='notebook_page'>
 +
<p>
 +
    IPTG stock: 50 ng/?l
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    July 26
 +
</p>
 +
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<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>
 +
    Glycerol stock and plasmid isolation
 +
</p>
 +
 +
<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>
 +
    Digestion and gel extraction
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LuxI-RBS-YFP (X/P): Concentration: 2.8 (to be repeated)
 +
    </li>
 +
</ul>
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    July 26
 +
</p>
 +
 +
<p>
 +
    Transformation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        BBa_C0012: LacI
 +
    </li>
 +
</div>
 +
<div id="page19" class='notebook_page'>
 +
 +
    <li>
 +
        BBa_I732100: LacI (2)
 +
    </li>
 +
</ul>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    July 27
 +
</p>
 +
 +
<p>
 +
    Glycerol stock and plasmid isolation
 +
</p>
 +
 +
<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>
 +
 +
<ul>
 +
    <li>
 +
        LasI-RBS-RFP: Concentration: 85 ng/µl
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion and gel extraction
 +
</p>
 +
 +
<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>
 +
 +
<ul>
 +
    <li>
 +
        LasI-RBS-RFP (X/P): Concentration: 8.9 ng/µl
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<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>
 +
    Transformation:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLsrA-LuxI-RBS-YFP
 +
    </li>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        BBa_B0422: RBS-CFP
 +
    </li>
 +
</ul>
 +
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    July 27
 +
</p>
 +
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LacI: 8 colonies seen: 1 colony picked
 +
    </li>
 +
    <li>
 +
        LacI (2): 2 colonies seen: 1 colony picked
 +
    </li>
 +
</ul>
 +
<p>
 +
    Plasmid isolation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LasR: Concentration: 95.8 ng/µl
 +
    </li>
 +
    <li>
 +
        Term-Term: Concentration: 120.1 ng/µl
 +
    </li>
 +
</ul>
 +
<p>
 +
    Toggle switch test
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        IPTG innoculation
 +
    </li>
 +
</div>
 +
<div id="page21" class='notebook_page'>
 +
 +
    <li>
 +
        TET innoculation
 +
    </li>
 +
    <li>
 +
        No innoculation
 +
    </li>
 +
</ul>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    July 28
 +
</p>
 +
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<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>
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    July 28
 +
</p>
 +
 +
<p>
 +
    Glycerol stock and plasmid isolation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LacI: Concentration: 73.9 ng/µl
 +
    </li>
 +
    <li>
 +
        LacI (2): Concentration: 109.8 ng/µl
 +
    </li>
 +
</ul>
 +
 +
</div>
 +
<div id="page22" class='notebook_page'>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    July 29
 +
</p>
 +
 +
<p>
 +
    Glycerol Stocks
 +
</p>
 +
 +
<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>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    July 30
 +
</p>
 +
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<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>
 +
 +
<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>
 +
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
</div>
 +
 +
<div id="page23" class='notebook_page'>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP
 +
    </li>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation and transformation:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Vector: pLasR
 +
    </li>
 +
    <li>
 +
        Insert: RBS-CFP
 +
    </li>
 +
    <li>
 +
        2 colonies shown: 1 colony picked
 +
    </li>
 +
</ul>
 +
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    July 30
 +
</p>
 +
 +
<p>
 +
    Experiment 7
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        150 ng/?l Tet
 +
    </li>
 +
    <li>
 +
        200 ng/?l Tet
 +
    </li>
 +
    <li>
 +
        300 ng/?l Tet
 +
    </li>
 +
    <li>
 +
        1mM IPTG
 +
    </li>
 +
    <li>
 +
        No Induction
 +
    </li>
 +
</ul>
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        RBS-GFP
 +
    </li>
 +
</div>
 +
<div id="page24" class='notebook_page'>
 +
 +
</ul>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    July 31
 +
</p>
 +
 +
<p>
 +
    Plasmid isolation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LuxS: Concentration: 78.1 ng/µl
 +
    </li>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP: Concentration: 48.7 ng/µl
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LuxS: (X/P); 516 bp
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR+LasI-RBS-RFP: Digested at XbaI and PstI; 1421 bp
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Test Digestion:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LuxS: digested at EcoRI and PstI
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-lasI-RBS-RFP: digested at EcoRI and PstI
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Colonies:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-lasI-RBS-RFP: 1 additional colony picked
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        LuxS: 1 additional colony picked
 +
    </li>
 +
</ul>
 +
 +
 +
</div>
 +
<div id="page25" class='notebook_page'>
 +
<p>
 +
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    July 31
 +
</p>
 +
 +
<p>
 +
    Plasmid Isolation:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        GFP: 137.5 ng/µl
 +
    </li>
 +
</ul>
 +
 +
 +
 +
<p>
 +
    August 1, 2012:
 +
</p>
 +
 +
<p>
 +
    Plasmid Isolation:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP: 61.2 ng/µl
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLasR-RBS-CFP: 49.2 ng/µl
 +
    </li>
 +
</ul>
 +
 +
 +
<p>
 +
    Digestion:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        GFP: Digested XbaI and PstI
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        LacI: Digested XbaI and PstI
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP: Digested XbaI and PstI
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLusR-RBS-CFP: Digested XbaI and PstI
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Gel Exraction:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        GFP: 8.5 ng/µl
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        LacI: 12.7 ng/µl
 +
    </li>
 +
</ul>
 +
 +
 +
</div>
 +
<div id="page26" class='notebook_page'>
 +
<p>
 +
    Colony:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLasR-RBS-CFP: 1 additional colony picked
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Transformation:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-lasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        BBa_J2309: pTetR-luxI
 +
    </li>
 +
</ul>
 +
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 1
 +
</p>
 +
 +
<p>
 +
    Ligation:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Vector: pTet; Insert: GFP
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        Vector: pTet; Insert: LacI
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        Vector: pLac; Insert: GFP
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Transformation:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pTet-GFP
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pTet-LacI
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLac-GFP
 +
    </li>
 +
</ul>
 +
 +
 +
 +
</div>
 +
<div id="page27" class='notebook_page'>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 2, 2012:
 +
</p>
 +
 +
<p>
 +
    Colonies:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP: about 5 colonies; 1 picked
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pTetR-LuxI: about 2 colonies; 1 picked
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Plasmid Isolation:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LuxS: Concentration – 40.5 ng/µl
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLasR-CFP: 95.8 ng/µl
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Digestion:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LasI: Digested at XbaI and PstI
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        LasR: Digested at XbaI and PstI
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        LuxS: Digested at XbaI and PstI
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLasR-RBS-CFP: Digested at XbaI and PstI
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Gel Extraction:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LasI: Concentration – 4.1 ng/µl
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        LasR: Concentration – 5.3 ng/µl
 +
    </li>
 +
 +
</div>
 +
<div id="page28" class='notebook_page'>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLasR-RBS-CFP – 13.4 ng/µl
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Ligation:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Vector: pLac
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        Insert: LasI
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        Transformation 2:1 – 3 colonies; 1 picked
 +
    </li>
 +
</ul>
 +
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 2
 +
</p>
 +
 +
<p>
 +
    Colonies:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pTet-GFP: 7 colonies; 1 picked
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pTet-LacI: 20 colonies; 1 picked
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLac-GFP: 20 colonies; 1 picked
 +
    </li>
 +
</ul>
 +
 +
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 3, 2012:
 +
</p>
 +
 +
<p>
 +
    Glycerol Stocks/Miniprep:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pTet-GFP: Concentration-142.2 ng/µl
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pTet-LacI: Concentration-67.2 ng/µl
 +
    </li>
 +
</div>
 +
<div id="page29" class='notebook_page'>
 +
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLac-GFP: Concentration-50.5 ng/µl
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Digestion:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pTet-GFP: Digested at SpeI and PstI
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pTet-LacI: Digested at SpeI and PstI
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLac-GFP: Digested at XbaI and PstI
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLac-TetR: Digested at XbaI and PstI
 +
    </li>
 +
</ul>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 3
 +
</p>
 +
 +
<p>
 +
    Ligation:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Vector: pTetR
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        Insert: LasR
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        Transformation 2:1 – 1 colony; 1 picked
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Ligation:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Vector: pLasR
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        Insert: RBS-CFP
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        Transformation: 2:1
 +
    </li>
 +
</ul>
 +
 +
</div>
 +
<div id="page30" class='notebook_page'>
 +
 +
<p>
 +
    Glycerol Stock:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP: Concentration – 45.1 ng/µl
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pTetR-LuxI: Concentration – 105.1 ng/µl
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Digestion:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP: Digested at XbaI and PstI
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Ligation:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Vector: pLac-luxR
 +
    </li>
 +
    <li>
 +
        Insert: pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Transformation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        BBa_K091109 (2010) LuxS
 +
    </li>
 +
    <li>
 +
        BBa_K091109 (2011) LuxS
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 4
 +
</p>
 +
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        BBa_K091109 (2010) LuxS: 70 colonies seen: 1 picked
 +
    </li>
 +
    <li>
 +
        BBa_K091109 (2011) LuxS: 100 colonies seen: 1 picked
 +
    </li>
 +
</ul>
 +
 +
 +
 +
 +
 +
</div>
 +
<div id="page31" class='notebook_page'>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 5
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
    <li>
 +
        pTetR-RBS-CFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Transformation (repeated)
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLasR-RBS-CFP
 +
    </li>
 +
    <li>
 +
       
 +
    </li>
 +
</ul>
 +
 +
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 6
 +
</p>
 +
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLasR-RBS-CFP: 10 colonies seen: 1 colony picked
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Plasmid isolation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LuxS: Concentration: 57.6
 +
    </li>
 +
 +
</div>
 +
<div id="page32" class='notebook_page'>
 +
    <li>
 +
        LuxS: Concentration: 76.4
 +
    </li>
 +
    <li>
 +
        pTetR-RBS-CFP: Concentration: 122.3
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Digestion and gel extraction (improperly prepared for gel)
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LuxS: Concentration: N/A
 +
    </li>
 +
    <li>
 +
        LuxS: Concentration: N/A
 +
    </li>
 +
    <li>
 +
        pTetR-RBS-CFP: Concentration: 21.9
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
    <li>
 +
        pTetR-RBS-CFP
 +
    </li>
 +
</ul>
 +
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 6, 2012:
 +
</p>
 +
 +
<p>
 +
    PCR:
 +
</p>
 +
 +
<p>
 +
    RFP-LVA created
 +
</p>
 +
 +
<p>
 +
    Experiments:
 +
</p>
 +
 +
<p>
 +
    Test mechanism shows TetR and LacI are medium strength repressors.
 +
 +
</div>
 +
<div id="page33" class='notebook_page'>
 +
 +
</p>
 +
 +
<p dir="ltr">
 +
    Toggle Experiment fails to show significant changes in fluorescence protein after 2 or 6 hours.
 +
</p>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 7
 +
</p>
 +
 +
 +
<p>
 +
    Glycerol Stock
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLasR-RBS-CFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Plasmid isolation
 +
</p>
 +
 +
<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>
 +
    Digestion and gel extraction
 +
</p>
 +
 +
<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>
 +
    Ligation
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Transformation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pSBIC3 chlora backbone
 +
    </li>
 +
    <li>
 +
        pLac-luxR-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 7, 2012
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock/Miniprep
 +
</p>
 +
 +
<p>
 +
    pTet-LacI
 +
</p>
 +
 +
<p>
 +
    pLac-GFP
 +
</p>
 +
 +
 +
<p>
 +
    Digestion &amp; Gel Extraction
 +
</p>
 +
 +
<p>
 +
    pTet-LacI: XbaI and PstI
 +
</p>
 +
 +
</div>
 +
<div id="page35" class='notebook_page'>
 +
 +
<p>
 +
    pLac-GFP: SpeI and PstI
 +
</p>
 +
 +
 +
<p>
 +
    Ligation/Transformation
 +
</p>
 +
 +
<p>
 +
    Vector: pLac-GFP
 +
</p>
 +
 +
<p>
 +
    Insert: pTet-LacI
 +
</p>
 +
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 8
 +
</p>
 +
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<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>
 +
    Repeat transformations from ligation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pyear-LuxS
 +
    </li>
 +
    <li>
 +
        pTetR-lasR-pLasR-RBS-CFP
 +
    </li>
 +
</ul>
 +
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 8, 2012
 +
</p>
 +
 +
 +
<p>
 +
    Transformations:
 +
 +
</div>
 +
<div id="page36" class='notebook_page'>
 +
</p>
 +
 +
<p>
 +
    Negative control had ~20 colonies
 +
</p>
 +
 +
<p>
 +
    2:1 sample had ~ 40 colonies, we picked 2
 +
</p>
 +
 +
<p>
 +
    Positive had too many colonies to count
 +
</p>
 +
 +
 +
<p>
 +
    PCR Reaction (Gradient)
 +
</p>
 +
 +
<p dir="ltr">
 +
    -55, 60, and 65 degrees
 +
</p>
 +
<p dir="ltr">
 +
    -55 degrees found to be most ideal
 +
</p>
 +
 +
<p>
 +
    Gel Extraction (of PCR)
 +
</p>
 +
 +
<p dir="ltr">
 +
    Poor results, no DNA extracted
 +
</p>
 +
 +
<p>
 +
    Plate test for toggle switch:
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 9
 +
</p>
 +
 +
<p>
 +
    Glycerol stocks
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        chlora backbone
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</div>
 +
<div id="page37" class='notebook_page'>
 +
 +
    <li>
 +
        pTetR-LasR-pLasR-RBS-CFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-Las-I
 +
    </li>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
</ul>
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        chlora backbone
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
    <li>
 +
        pTetR-LasR-pLasR-RBS-CFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Term-Term (S/P)
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        chlora backbone (E/P)
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-pLuxR-LasI-RBS-RFP (E/P)
 +
    </li>
 +
    <li>
 +
        pTetR-LasR-pLasR-RBS-CFP (E/P)
 +
    </li>
 +
</ul>
 +
<p>
 +
    Gel Extraction
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Term-Term: Concentration: 44.3
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        chlora backbone: Concentration: 332.9
 +
    </li>
 +
</div>
 +
<div id="page38" class='notebook_page'>
 +
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-pLuxR-LasI-RBS-RFP: bad results
 +
    </li>
 +
    <li>
 +
        pTetR-LasR-pLasR-RBS-CFP: bad results
 +
    </li>
 +
</ul>
 +
<p>
 +
    Innoculation: pLac-LuxR-pLuxR-LasI-RBS-RFP
 +
</p>
 +
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 9, 2012
 +
</p>
 +
 +
<p>
 +
    Glycerol stock/miniprep:
 +
</p>
 +
 +
<p>
 +
    pLac-GFP-ST-pTet-LacI
 +
</p>
 +
 +
 +
<p>
 +
    PCR Reaction
 +
</p>
 +
 +
<p>
 +
    RFP-LVA
 +
</p>
 +
 +
 +
<p>
 +
    Experiment: Repressor Test Mechanism (LacI)
 +
</p>
 +
 +
<p>
 +
    1 - Induced 10microliters of 10mM IPTG
 +
</p>
 +
 +
<p>
 +
    1 - No induction
 +
</p>
 +
 +
 +
<p>
 +
    Digestion and Gel Extraction:
 +
</p>
 +
 +
<p>
 +
    RFP-LVA: XbaI and PstI
 +
</p>
 +
 +
<p>
 +
    Ampicillin Backbone: SpeI and PstI
 +
</p>
 +
 +
 +
<p>
 +
 +
</div>
 +
<div id="page39" class='notebook_page'>
 +
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 10
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Plasmid isolation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-pLuxR-LasI-RBS-RFP: Concentration: 125.1
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-pLuxR-LasI-RBS-RFP (E/P)
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<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>
 +
    Transformation:
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Term-Term-pLasR-RBS-CFP
 +
    </li>
 +
    <li>
 +
        Term-Term-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</div>
 +
<div id="page40" class='notebook_page'>
 +
 +
</ul>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 10, 2012
 +
</p>
 +
 +
<p>
 +
    Ligation/Transformation
 +
</p>
 +
 +
<p>
 +
    RFP-LVA into Ampicillin backbone
 +
</p>
 +
 +
 +
<p>
 +
    Inoculations:
 +
</p>
 +
 +
<p>
 +
    Toggle
 +
</p>
 +
 +
<p>
 +
    pLas
 +
</p>
 +
 +
<p>
 +
    LacI
 +
</p>
 +
 +
<p>
 +
    LuxR
 +
</p>
 +
 +
<p>
 +
    pTet
 +
</p>
 +
 +
<p>
 +
    pLux
 +
</p>
 +
 +
<p>
 +
    LuxI
 +
</p>
 +
 +
 +
<p>
 +
    August 11, 2012
 +
</p>
 +
 +
<p>
 +
    Plasmid isolation
 +
</p>
 +
 +
<p>
 +
    Toggle
 +
</p>
 +
 +
<p>
 +
    pLas
 +
</div>
 +
<div id="page41" class='notebook_page'>
 +
 +
</p>
 +
 +
<p>
 +
    LacI
 +
</p>
 +
 +
<p>
 +
    LuxR
 +
</p>
 +
 +
<p>
 +
    pTet
 +
</p>
 +
 +
<p>
 +
    pLux
 +
</p>
 +
 +
<p>
 +
    LuxI
 +
</p>
 +
 +
 +
<p>
 +
    Digestion/Extraction
 +
</p>
 +
 +
<p>
 +
    LuxR: X and P
 +
</p>
 +
 +
<p>
 +
    pLux: S and P
 +
</p>
 +
 +
<p>
 +
    LuxI: S and P
 +
</p>
 +
 +
<p>
 +
    LasR: X and P
 +
</p>
 +
 +
<p>
 +
    Double Terminators (DT): X and P
 +
</p>
 +
 +
<p>
 +
    LasI: X and P
 +
</p>
 +
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 11
 +
</p>
 +
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<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>
 +
 +
 +
</div>
 +
<div id="page42" class='notebook_page'>
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 12
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock and Plasmid isolation
 +
</p>
 +
 +
<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>
 +
 +
<ul>
 +
    <li>
 +
        pyear-LuxS
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion
 +
</p>
 +
 +
<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>
 +
 +
<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>
 +
 +
<ul>
 +
    <li>
 +
        Vector1: pTetR-LasR
 +
    </li>
 +
    <li>
 +
        Insert1: Term-Term-pLasR-RBS-CFP
 +
    </li>
 +
 +
</div>
 +
<div id="page43" class='notebook_page'>
 +
 +
    <li>
 +
        Vector2: pLac-LuxR
 +
    </li>
 +
    <li>
 +
        Insert 2: Term-Term-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 13
 +
</p>
 +
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-LasI
 +
    </li>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 13, 2012
 +
</p>
 +
 +
<p>
 +
    Miniprep/Glycerol Stock
 +
</p>
 +
 +
<p>
 +
    RFP-LVA
 +
</p>
 +
 +
 +
<p>
 +
    Digestions/Extraction
 +
</p>
 +
 +
<p>
 +
    pLas: S and P
 +
</p>
 +
 +
<p>
 +
    LacI: X and P
 +
</p>
 +
 +
<p>
 +
    RFP-LVA: E and P
 +
</p>
 +
 +
 +
<p>
 +
    Inoculations
 +
 +
</div>
 +
<div id="page43" class='notebook_page'>
 +
 +
</p>
 +
 +
<p>
 +
    Chlorabackbone
 +
</p>
 +
 +
<p>
 +
    RFP-LVA
 +
</p>
 +
 +
<p>
 +
    DT
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 14
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock and Plasmid isolation
 +
</p>
 +
 +
<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>
 +
 +
<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>
 +
 +
<p>
 +
    Parts Recieved
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        BBa_K091159
 +
    </li>
 +
    <li>
 +
        BBa_K091002
 +
    </li>
 +
    <li>
 +
        BBa_I13453
 +
    </li>
 +
    <li>
 +
        BBa_B0034
 +
    </li>
 +
</div>
 +
<div id="page44" class='notebook_page'>
 +
 +
</ul>
 +
<p>
 +
    Streaking
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        BBa_K091159 pBad-LsrA
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation repeat
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 14, 2012
 +
</p>
 +
 +
<p>
 +
    Miniprep/Glycerol Stock
 +
</p>
 +
 +
<p>
 +
    Chlorabackbone
 +
</p>
 +
 +
<p>
 +
    DT
 +
</p>
 +
 +
 +
<p>
 +
    Digestion/Extraction:
 +
</p>
 +
 +
<p>
 +
    DT: X and P
 +
</p>
 +
 +
<p>
 +
    Chlorabackbone: E and P
 +
</p>
 +
 +
<p>
 +
    Toggle: E and P
 +
 +
</div>
 +
<div id="page45" class='notebook_page'>
 +
 +
</p>
 +
 +
 +
<p>
 +
    Ligation/Transformation:
 +
</p>
 +
 +
<p>
 +
    DT into LuxR
 +
</p>
 +
 +
<p>
 +
    TetR into pLux
 +
</p>
 +
 +
<p>
 +
    LasI into pTet
 +
</p>
 +
 +
<p>
 +
    LasR into DT
 +
</p>
 +
 +
<p>
 +
    pLas into LacI
 +
</p>
 +
 +
<p dir="ltr">
 +
    pLac into LuxI
 +
</p>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 15
 +
</p>
 +
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
    <li>
 +
        pLac-LasI
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        pTetR-LasR-Term-Term-pLasR-RBS-CFP
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-Term-Term-pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pBad-LsrA: 30 colonies seen: 1 picked
 +
    </li>
 +
</ul>
 +
 +
</div>
 +
<div id="page46" class='notebook_page'>
 +
 +
<p>
 +
    Testing
 +
</p>
 +
 +
<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>
 +
    Streaking
 +
</p>
 +
 +
<p>
 +
    BBa_K091109
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        SP3011&gt;1D LuxS
 +
    </li>
 +
    <li>
 +
        SP4011&gt;1D LuxS
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 15, 2012
 +
</p>
 +
 +
<p>
 +
    Transformation
 +
</p>
 +
 +
</div>
 +
<div id="page47" class='notebook_page'>
 +
<p>
 +
    LuxR-DT: 15 colonies, picked 1 colony
 +
</p>
 +
 +
<p>
 +
    pLux-TetR: 35 colonies, picked 1
 +
</p>
 +
 +
<p>
 +
    pTet-LasI: Negative control growth; unsuccessful
 +
</p>
 +
 +
<p>
 +
    LasR-DT: 25 colonies, picked 1
 +
</p>
 +
 +
<p>
 +
    pLas-LacI: 0 growth
 +
</p>
 +
 +
<p>
 +
    pLac-LuxI: 15 colonies, picked 1
 +
</p>
 +
 +
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<p>
 +
    LasI into pTet
 +
</p>
 +
 +
 +
<p>
 +
    Transformation
 +
</p>
 +
 +
<p>
 +
    pTet-LasI
 +
</p>
 +
 +
<p>
 +
    pLas-LacI
 +
</p>
 +
 +
 +
<p>
 +
    Inoculations
 +
</p>
 +
 +
<p>
 +
    Chlorabackbone
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 16
 +
</p>
 +
 +
<p>
 +
    Colonies
 +
</div>
 +
<div id="page48" class='notebook_page'>
 +
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pTetR-LuxI:
 +
    </li>
 +
    <li>
 +
        pLac-LasI
 +
    </li>
 +
</ul>
 +
 +
<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>
 +
 +
<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>
 +
 +
<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 id="page49" class='notebook_page'>
 +
 +
<p>
 +
    Testing
 +
</p>
 +
 +
<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'>
 +
 +
        <li>
 +
            100/400
 +
        </li>
 +
        <li>
 +
            100/600
 +
        </li>
 +
        <li>
 +
            Results: After 6 and 24 hours: no considerable difference
 +
        </li>
 +
    </ul>
 +
</ul>
 +
<p>
 +
    Digestion
 +
</p>
 +
 +
<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>
 +
    Gel Extraction
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pBad-lsrK: Concentration: 53.8
 +
    </li>
 +
    <li>
 +
        pLsrA-LuxI-RBS-YFP: Concentration: 20.0
 +
    </li>
 +
</ul>
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Term-term
 +
    </li>
 +
</ul>
 +
 +
 +
</div>
 +
<div id="page51" class='notebook_page'>
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 16, 2012
 +
</p>
 +
 +
<p>
 +
    Miniprep/Glycerol Stock
 +
</p>
 +
 +
<p>
 +
    LuxR-DT
 +
</p>
 +
 +
<p>
 +
    pLux-TetR
 +
</p>
 +
 +
<p>
 +
    LasR-DT
 +
</p>
 +
 +
<p>
 +
    pLac-LacI
 +
</p>
 +
 +
<p>
 +
    Chlorabackbone
 +
</p>
 +
 +
<p>
 +
    Transformation Results
 +
</p>
 +
 +
<p>
 +
    Negative controls had too much growth; results thrown out
 +
</p>
 +
 +
 +
<p>
 +
    Digestion/Extraction
 +
</p>
 +
 +
<p>
 +
    LuxR-DT: S and P
 +
</p>
 +
 +
<p>
 +
    pLux-TetR: X and P
 +
</p>
 +
 +
<p>
 +
    Chlorabackbone: E and P
 +
</p>
 +
 +
<p>
 +
    Ligations/Transformation
 +
</p>
 +
 +
<p>
 +
    LuxR-DT and pLux-TetR
 +
</div>
 +
<div id="page52" class='notebook_page'>
 +
 +
</p>
 +
 +
<p>
 +
    Chlorabackbone with Toggle Switch
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 17
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Population 3 w/ Term-Term
 +
    </li>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
    <li>
 +
        LuxS
 +
    </li>
 +
</ul>
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<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>
 +
 +
<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'>
 +
 +
    <li>
 +
        Term-Term: Concentration: gel not used
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<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>
 +
    Transformation
 +
</p>
 +
 +
<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>
 +
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 17, 2012
 +
</p>
 +
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<p>
 +
    pLas with LacI
 +
</p>
 +
 +
<p>
 +
    pTet with LasI
 +
</p>
 +
 +
 +
<p>
 +
    Inoculations
 +
</p>
 +
 +
<p>
 +
    pLas
 +
</p>
 +
 +
<p>
 +
    LacI
 +
</p>
 +
 +
<p>
 +
    pTet
 +
</p>
 +
 +
<p>
 +
    pLac
 +
</p>
 +
 +
<p>
 +
    GFP
 +
</p>
 +
 +
<p>
 +
    RFP-LVA
 +
</p>
 +
 +
<p>
 +
    LasI
 +
</p>
 +
 +
 +
<p>
 +
    Transformation
 +
</p>
 +
 +
<p>
 +
    Bba_K081014 Kit Plate 3 Well 12O Resistance AK; RFP
 +
</p>
 +
 +
 +
</div>
 +
<div id="page55" class='notebook_page'>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 18
 +
</p>
 +
 +
<p>
 +
    Innoculations
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pTetR-LuxI
 +
    </li>
 +
    <li>
 +
        pLac-lasI
 +
    </li>
 +
    <li>
 +
        pop 2 w/TT (oscillator)
 +
    </li>
 +
    <li>
 +
        pop 3 w/TT (oscillator)
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 18, 2012
 +
</p>
 +
 +
<p>
 +
    Colony Picking
 +
</p>
 +
 +
<p>
 +
    LuxR-DT-pLux-TetR: 30 colonies, 1 picked
 +
</p>
 +
 +
<p>
 +
    Toggle Switch in Chlorabackbone: 4 colonies, 1 picked
 +
</p>
 +
 +
<p>
 +
    pLas with LacI: negative control growth; results thrown out
 +
</p>
 +
 +
<p>
 +
    pTet with LasI: negative control growth; results thrown out
 +
</p>
 +
 +
<p dir="ltr">
 +
    RFP: 20 colonies, 1 picked
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock
 +
</p>
 +
 +
</div>
 +
<div id="page56" class='notebook_page'>
 +
 +
<p>
 +
    LuxR-DT-pLux-TetR
 +
</p>
 +
 +
 +
<p>
 +
    Minipreps
 +
</p>
 +
 +
<p>
 +
    LuxR-DT-pLux-TetR
 +
</p>
 +
 +
<p>
 +
    pLas
 +
</p>
 +
 +
<p>
 +
    LacI
 +
</p>
 +
 +
<p>
 +
    pTet
 +
</p>
 +
 +
<p>
 +
    pLac
 +
</p>
 +
 +
<p>
 +
    GFP
 +
</p>
 +
 +
<p>
 +
    LasI
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
</div>
 +
<div id="page57" class='notebook_page'>
 +
 +
<p>
 +
    August 19
 +
</p>
 +
 +
<p>
 +
    Glycerol Stocks
 +
</p>
 +
 +
<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>
 +
 +
<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'>
 +
 +
    <li>
 +
        pop2 and pop3 75:1 - a visual increase of both RFP and CFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<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>
 +
    Digestion and gel extraction
 +
</p>
 +
 +
<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>
 +
    Ligation
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 19, 2012
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock/Plasmid Isolation
 +
</p>
 +
 +
<p>
 +
    RFP
 +
</p>
 +
 +
<p>
 +
    Toggle Switch in Chlorabackbone
 +
</p>
 +
 +
 +
<p>
 +
    Inoculation
 +
</p>
 +
 +
<p>
 +
    RFP-LVA
 +
</p>
 +
 +
 +
<p>
 +
    Digestion/Extraction
 +
</p>
 +
 +
<p>
 +
    pLac: S and P
 +
</p>
 +
 +
<p>
 +
    LuxR-DT-pLux-TetR: X and P
 +
</p>
 +
 +
<p>
 +
    pTet: S and P
 +
</p>
 +
 +
<p>
 +
    LasI: X and P
 +
</p>
 +
 +
<p>
 +
    LasR-DT: X and P
 +
</p>
 +
 +
</div>
 +
<div id="page61" class='notebook_page'>
 +
 +
<p>
 +
    LacI: S and P
 +
</p>
 +
 +
<p>
 +
    RFP: X and P
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 20
 +
</p>
 +
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<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>
 +
    Innoculation
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 20, 2012
 +
</p>
 +
 +
<p>
 +
    Ligations/Transformations
 +
</p>
 +
 +
<p>
 +
    LuxR-DT-pLux-TetR into pLac
 +
</p>
 +
 +
<p>
 +
    LasI into pTet
 +
</p>
 +
 +
<p>
 +
    LasR-DT into pTet
 +
</p>
 +
 +
<p>
 +
    RFP into LacI
 +
</p>
 +
 +
 +
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 21
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock and plasmid isolation
 +
</p>
 +
 +
<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 id="page63" class='notebook_page'>
 +
 +
 +
    <li>
 +
        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>
 +
 +
<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>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pyear-LuxS
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 21, 2012
 +
</p>
 +
 +
<p>
 +
    Colony Pickings
 +
</p>
 +
 +
<p>
 +
    pLac-LuxR-DT-pLux-TetR: 24 colonies, 1 picked
 +
</p>
 +
 +
<p>
 +
    pTet-LasI: 30 colonies, 3 picked
 +
</p>
 +
 +
<p>
 +
    pTet-LasR-DT: 100 colonies, 2 picked
 +
</p>
 +
 +
<p>
 +
    LacI-RFP: 24 colonies, 2 picked
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 22
 +
</p>
 +
 +
<p>
 +
    Digestion (test)
 +
</p>
 +
 +
<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>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        population 1 (oscillator)
 +
    </li>
 +
</ul>
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<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>
 +
    Transformation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-LuxR
 +
    </li>
 +
</ul>
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
 +
</div>
 +
<div id="page66" class='notebook_page'>
 +
 +
<p>
 +
    Population Testing Prep (pop2, pop3, pop2+pop3)
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 22, 2012
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock/Miniprep
 +
</p>
 +
 +
<p>
 +
    pLac-LuxR-DT-pLux-TetR
 +
</p>
 +
 +
<p>
 +
    pTet-LasI
 +
</p>
 +
 +
<p>
 +
    pTet-LasR-DT
 +
</p>
 +
 +
<p>
 +
    LacI-RFP
 +
</p>
 +
 +
 +
<p>
 +
    Digestion/Extraction:
 +
</p>
 +
 +
<p>
 +
    pLac-LuxR-DT-pLux-TetR: S and P
 +
</p>
 +
 +
</div>
 +
<div id="page67" class='notebook_page'>
 +
 +
<p>
 +
    GFP: X and P
 +
</p>
 +
 +
<p>
 +
    LacI-RFP: S and P
 +
</p>
 +
 +
<p>
 +
    pLac-LuxI: X and P
 +
</p>
 +
 +
<p>
 +
    Test Digestions
 +
</p>
 +
 +
<p dir="ltr">
 +
    pTet-LasI: E and P
 +
</p>
 +
<p>
 +
    pTet-LasR-DT: E and P
 +
</p>
 +
 +
 +
<p>
 +
    Ligations/Transformations
 +
</p>
 +
 +
<p>
 +
    GFP into pLac-LuxR-DT-pLux-TetR
 +
</p>
 +
 +
<p>
 +
    pLac-LuxI into LacI-RFP
 +
</p>
 +
 +
 +
<p>
 +
    Inoculations
 +
</p>
 +
 +
<p>
 +
    RFP-LVA
 +
</p>
 +
 +
<p>
 +
    pTet-GFP
 +
</p>
 +
 +
<p>
 +
    RBS-YFP
 +
</p>
 +
 +
 +
<p>
 +
    August 22
 +
</p>
 +
 +
<p>
 +
    Digestion (test)
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        population 2 (oscillator)
 +
    </li>
 +
</div>
 +
<div id="page68" class='notebook_page'>
 +
 +
    <ul>
 +
        <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>
 +
 +
<ul>
 +
    <li>
 +
        population 1 (oscillator)
 +
    </li>
 +
</ul>
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<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>
 +
    Transformation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-LuxR
 +
    </li>
 +
</ul>
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
</div>
 +
<div id="page69" class='notebook_page'>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Population Testing Prep (pop2, pop3, pop2+pop3)
 +
</p>
 +
 +
<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>
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 23, 2012
 +
</p>
 +
 +
 +
<p>
 +
    Colony Picking
 +
</p>
 +
 +
<p>
 +
    LacI-RFP-ST-pLac-LuxI; 40 colonies, 1 picked
 +
</p>
 +
 +
<p>
 +
    LacI-RFP-ST-pLac-LuxI; 30 colonies, 1 picked
 +
</p>
 +
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 23
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock
 +
</p>
 +
 +
 +
</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>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLasR-TetO-RBS-CFP: Concentration: 113.7
 +
    </li>
 +
    <li>
 +
        pLasR-TetO-RBS-CFP: Concentration: 112.0
 +
    </li>
 +
</ul>
 +
 +
<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>
 +
    Colonies picked
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-LuxR
 +
    </li>
 +
</ul>
 +
<p>
 +
    Population Testing from 8/22 prep after 24 hours
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        500 microliters from test tube of populations put in microcentrifuge tubes
 +
    </li>
 +
</ul>
 +
 +
<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 id="page71" class='notebook_page'>
 +
        <li>
 +
            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>
 +
    Digestion
 +
</p>
 +
 +
<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>
 +
    Innoculation
 +
</p>
 +
 +
<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>
 +
    Ligation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Vector: pBad-TetR-TT
 +
    </li>
 +
    <li>
 +
        Insert: plasR-TetO-RBS-CFP
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 24, 2012
 +
</p>
 +
 +
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<p>
 +
    LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
 +
 +
<p>
 +
    Digestion/Extraction
 +
</p>
 +
 +
<p>
 +
    pTet-LasR-DT
 +
</p>
 +
 +
<p>
 +
    pTet-LasI
 +
</p>
 +
 +
<p>
 +
    LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
 +
<p>
 +
    pLas
 +
</p>
 +
 +
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<p>
 +
    pTet-LasI into pLac-LuxR-DT-pLux-TetR-GFP-ST
 +
</p>
 +
 +
 +
</div>
 +
<div id="page74" class='notebook_page'>
 +
<p>
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
</div>
 +
    August 24
 +
</p>
 +
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pluxR-LasI-RBS-RFP: Concentration: 42.6
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP (X/P)
 +
    </li>
 +
    <li>
 +
        pLuxR-araC-lasR-TT (X/P)
 +
    </li>
 +
</ul>
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-LasI-RBS-RFP
 +
    </li>
 +
    <li>
 +
        pLuxR-araC-LasR-TT
 +
    </li>
 +
    <li>
 +
        pLsrA-LuxI-YFP
 +
    </li>
 +
</ul>
 +
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 25, 2012
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock/Plasmid Isolation
 +
</p>
 +
 +
<p>
 +
    LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
 +
 +
<p>
 +
    Digestion/Extraction
 +
</p>
 +
 +
</div>
 +
<div id="page75" class='notebook_page'>
 +
<p>
 +
<p>
 +
    LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
 +
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<p>
 +
    LacI-RFP-ST-pLac-LuxI into pLas
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 25
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock
 +
</p>
 +
 +
<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>
 +
 +
<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>
 +
    Digestion and Gel Extraction
 +
</p>
 +
 +
</div>
 +
<div id="page76" class='notebook_page'>
 +
<p>
 +
 +
<ul>
 +
    <li>
 +
        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>
 +
    Ligation
 +
</p>
 +
 +
<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>
 +
    Primer Dilution
 +
</p>
 +
 +
<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>
 +
    Colony PCR
 +
</p>
 +
 +
</div>
 +
<div id="page77" class='notebook_page'>
 +
<p>
 +
<ul>
 +
    <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>
 +
    Innoculation
 +
</p>
 +
 +
<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>
 +
    August 26, 2012
 +
</p>
 +
 +
<p>
 +
    Colony Picking
 +
</p>
 +
 +
<p>
 +
    pLas-LacI-RFP-ST-pLac-LuxI; 50 colonies, 1 picked
 +
</p>
 +
 +
 +
<p>
 +
    Inoculation
 +
</p>
 +
 +
<p>
 +
    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP
 +
</p>
 +
 +
<p>
 +
    pLas-LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 26
 +
</p>
 +
 +
<p>
 +
    Colonies Picked
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-araC-LasR-TT: 1 colony picked
 +
    </li>
 +
</ul>
 +
<p>
 +
    Population testing
 +
</p>
 +
 +
<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 id="page79" class='notebook_page'>
 +
 +
    </ul>
 +
</ul>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 27, 2012
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock/Plasmid Isolation
 +
</p>
 +
 +
<p>
 +
    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP
 +
</p>
 +
 +
<p>
 +
    pLas-LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
 +
 +
<p>
 +
    PCR
 +
</p>
 +
 +
<p>
 +
    Add second LacO site to pLac
 +
</p>
 +
 +
 +
<p>
 +
    Digestion/Extraction
 +
</p>
 +
 +
<p>
 +
    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP
 +
</p>
 +
 +
<p>
 +
    pLas-LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 27
 +
</p>
 +
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-araC-LaR-TT: Concentration: 164.3
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion and Gel extraction
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLuxR-araC-LaR-TT (X/P): Concentration: 55.1
 +
    </li>
 +
</ul>
 +
</div>
 +
<div id="page80" class='notebook_page'>
 +
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
<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>
 +
    Ligation
 +
</p>
 +
 +
<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 id="page81" class='notebook_page'>
 +
    <ul>
 +
        <li>
 +
            0:1 5 colonies seen
 +
        </li>
 +
        <li>
 +
            2:1 25 colonies seen
 +
        </li>
 +
    </ul>
 +
</ul>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 28, 2012
 +
</p>
 +
 +
<p>
 +
    Primer Dilutions
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 28
 +
</p>
 +
 +
<p>
 +
    Colony PCR
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pBad-tetR-TT-pLasR-TetO-RBS-CFP
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-TT- pLuXR-araC-lasR-TT
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        Primer 1: I3.24.SP1
 +
    </li>
 +
    <li>
 +
        Primer 2: I3.PSB1A2.R
 +
    </li>
 +
</ul>
 +
<p>
 +
    Population Testing
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        200ul medium, 200ul induction, 4ul population, 24 hour incubation
 +
    </li>
 +
</ul>
 +
 +
<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>
 +
    Ligation
 +
</p>
 +
 +
<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>
 +
    Digestion and Gel Extraction
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pBad-TetR-TT (S/P): Concentration: 88.3
 +
    </li>
 +
</ul>
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 29, 2012
 +
</p>
 +
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<p>
 +
    pLas-LacI-RFP-ST-pLac-LuxI into pTet-LasR-DT
 +
</p>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 29
 +
</p>
 +
 +
<p>
 +
    Population Test Prep
 +
</p>
 +
 +
<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 id="page84" class='notebook_page'>
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 30
 +
</p>
 +
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<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>
 +
    Colony PCR
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-TT-pLuXR-araC-lasR-TT (X2)
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        Primer 1: I3.24.SP1
 +
    </li>
 +
    <li>
 +
        Primer 2: I3.PSB1A2.R
 +
    </li>
 +
</ul>
 +
<p>
 +
    Colonies picked
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-TT-pLuXR-araC-lasR-TT (X2)
 +
    </li>
 +
</ul>
 +
 +
</div>
 +
<div id="page85" class='notebook_page'>
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    August 31, 2012
 +
</p>
 +
 +
<p>
 +
    Colony picking
 +
</p>
 +
 +
<p>
 +
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI; 50 colonies, 1 picked
 +
</p>
 +
 +
 +
<p>
 +
    PCR Reaction
 +
</p>
 +
 +
<p>
 +
    Add LacO binding site to pLac; 65.2 degrees found to be optimal temperature for elongation
 +
</p>
 +
 +
 +
<p>
 +
    Transformation
 +
</p>
 +
 +
<p>
 +
    pLac-LacO
 +
</p>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    August 31
 +
</p>
 +
 +
 +
<p>
 +
    Glycerol Stock and Plasmid Isolation
 +
</p>
 +
 +
<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>
 +
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-TT-pLuXR-araC-lasR-TT (S/P)
 +
    </li>
 +
</div>
 +
<div id="page86" class='notebook_page'>
 +
    <li>
 +
        pLac-LuxR-TT-pLuXR-araC-lasR-TT (S/P)
 +
    </li>
 +
</ul>
 +
<p>
 +
    Colony PCR
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pBad-TetR-TT-pLasR-TetO-CFP (X2)
 +
    </li>
 +
</ul>
 +
 +
<ul>
 +
    <li>
 +
        Primer 1: I3.24.SP1
 +
    </li>
 +
    <li>
 +
        Primer 2: I3.PSB1A2.R
 +
    </li>
 +
</ul>
 +
<p>
 +
    Population Test Results
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    September 1, 2012
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock/Plasmid Isolation
 +
</p>
 +
 +
<p>
 +
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
</div>
 +
<div id="page87" class='notebook_page'>
 +
 +
 +
<p>
 +
    Colony Picking
 +
</p>
 +
 +
<p>
 +
    pLac-LacO
 +
</p>
 +
 +
 +
<p>
 +
    Inoculations
 +
</p>
 +
 +
<p>
 +
    pLac-TetR
 +
</p>
 +
 +
<p>
 +
    pLac-TetR-RBS-RFP
 +
</p>
 +
 +
<p>
 +
    pTet-LacI
 +
</p>
 +
 +
<p>
 +
    pTet-RBS-GFP-T
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    September 1
 +
</p>
 +
 +
<p>
 +
    Ligations
 +
</p>
 +
 +
<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 id="page88" class='notebook_page'>
 +
    <li>
 +
        Insert3: pLsrA-LuxI-RBS-YFP
 +
    </li>
 +
    <li>
 +
        Vector4: pLuxR
 +
    </li>
 +
    <li>
 +
        Insert4: TetR
 +
    </li>
 +
</ul>
 +
<p>
 +
    Transformation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        BBa_C0012: LacI
 +
    </li>
 +
    <li>
 +
        BBa_K091002: LsrK
 +
    </li>
 +
</ul>
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLacI-LuxR-TT-pLuxR-LasI-RBS-RFP (pop2 oscillator)
 +
    </li>
 +
</ul>
 +
 +
 +
</div>
 +
<div id="page89" class='notebook_page'>
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    September 2, 2012
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock/Plasmid Isolation
 +
</p>
 +
 +
<p>
 +
    pLac-LacO
 +
</p>
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    September 3, 2012
 +
</p>
 +
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<p>
 +
    pLac-TetR
 +
</p>
 +
 +
<p>
 +
    pLac-TetR-RBS-RFP
 +
</p>
 +
 +
<p>
 +
    pTet-LacI
 +
</p>
 +
 +
<p>
 +
    pTet-RBS-GFP-T
 +
</p>
 +
 +
 +
<p>
 +
    Digestion/Extraction
 +
</p>
 +
 +
<p dir="ltr">
 +
    pLac-TetR
 +
</p>
 +
<p>
 +
    pLac-TetR-RBS-RFP
 +
</p>
 +
 +
<p>
 +
    pTet-LacI
 +
</p>
 +
 +
<p>
 +
    pTet-RBS-GFP-T
 +
</p>
 +
 +
</div>
 +
<div id="page90" class='notebook_page'>
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    September 3
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac-LuxR-TT-pLuxR-araC-LasR-TT
 +
    </li>
 +
    <li>
 +
        pBad-TetR-TT-pLas-TetO-CFP
 +
    </li>
 +
</ul>
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<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>
 +
    Digestion
 +
</p>
 +
 +
<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>
 +
    Innoculation
 +
</p>
 +
 +
<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>
 +
    Colony PCR for I3-98-2 and I4-1-2-
 +
</p>
 +
 +
<p>
 +
    Digestion
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        TT-pLuxR-LasI-RFP (X/P)
 +
    </li>
 +
    <li>
 +
        pLac-LuxR (S/P)
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    September 4, 2012
 +
</p>
 +
 +
<p>
 +
    Inoculations
 +
</p>
 +
 +
<p>
 +
    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI (Atc 10 microliters)
 +
</p>
 +
 +
<p>
 +
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI (IPTG 10 microliters)
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    September 4
 +
</p>
 +
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pBad-TetR-TT-pLasR-TetO-CFP: Concentration: 190.4
 +
    </li>
 +
</ul>
 +
</div>
 +
<div id="page92" class='notebook_page'>
 +
<p>
 +
    Digestion
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pBad-TetR-TT-pLasR-TetO-CFP (X/P): Concentration: 20.5
 +
    </li>
 +
</ul>
 +
<p>
 +
    Colonies picked from I4-1-2
 +
</p>
 +
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    September 5, 2012
 +
</p>
 +
 +
<p>
 +
    Inoculations
 +
</p>
 +
 +
<p dir="ltr">
 +
    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI
 +
</p>
 +
<p>
 +
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
 +
<p>
 +
    1 with Atc
 +
</p>
 +
 +
<p>
 +
    1 with IPTG
 +
</p>
 +
 +
<p>
 +
    1 with No induction
 +
</p>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    September 5
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock and Plasmid Isolation
 +
</p>
 +
 +
<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>
 +
    Digestion and Gel Extraction
 +
</p>
 +
 +
<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>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac
 +
    </li>
 +
    <li>
 +
        LasI-RFP
 +
    </li>
 +
</ul>
 +
 +
</div>
 +
<div id="page94" class='notebook_page'>
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    September 6, 2012
 +
</p>
 +
 +
<p>
 +
    Sequencing
 +
</p>
 +
 +
 +
<p>
 +
    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI
 +
</p>
 +
 +
<p>
 +
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
 +
<p>
 +
    pLac-LacO
 +
</p>
 +
 +
<p>
 +
    RFP-LVA
 +
</p>
 +
 +
 +
<p>
 +
    Inoculations
 +
</p>
 +
 +
<p>
 +
    Chlorabackbone
 +
</p>
 +
 +
<p dir="ltr">
 +
    pLac-LuxR-DT-pLux-TetR-GFP-ST-pTet-LasI
 +
</p>
 +
<p>
 +
    pTet-LasR-DT-pLas-LacI-RFP-ST-pLac-LuxI
 +
</p>
 +
 +
<p>
 +
    1 with Atc
 +
</p>
 +
 +
<p>
 +
    1 with IPTG
 +
</p>
 +
 +
<p>
 +
    1 with No induction
 +
</p>
 +
 +
 +
<p>
 +
    Digestion/Extraction
 +
</p>
 +
 +
<p>
 +
    Chlorabackbone
 +
</p>
 +
 +
 +
</div>
 +
<div id="page95" class='notebook_page'>
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    September 6
 +
</p>
 +
 +
<p>
 +
    Plasmid Isolation and Glycerol Stock
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac
 +
    </li>
 +
    <li>
 +
        LasI-RFP
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-TT-pLuxR-araC-LasR-TT
 +
    </li>
 +
</ul>
 +
<p>
 +
    Digestion and Gel Extraction
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac
 +
    </li>
 +
    <li>
 +
        LasI-RFP
 +
    </li>
 +
    <li>
 +
        pLac-LuxR-TT-pLuxR-araC-LasR-TT
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    September 7, 2012
 +
</p>
 +
 +
<p>
 +
    Inoculation
 +
</p>
 +
 +
<p>
 +
    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP
 +
</p>
 +
 +
<p>
 +
    pLac-RBS-GFP-T
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    September 7
 +
</p>
 +
 +
<p>
 +
    Test Digestions
 +
</p>
 +
 +
<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>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Term-Term
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    September 8, 2012
 +
</p>
 +
 +
<p>
 +
    Time-Lapse Experiment to observe toggle switch
 +
</p>
 +
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<p dir="ltr">
 +
    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP
 +
</p>
 +
<p>
 +
    pLac-RBS-GFP-T
 +
</p>
 +
 +
<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>
 +
</div>
 +
<div id="page99" class='notebook_page'>
 +
<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>
 +
<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>
 +
 +
<p>
 +
    Ligations
 +
</p>
 +
 +
<p>
 +
    pLac-TetR into Chlora
 +
</p>
 +
 +
<p>
 +
    pLac-RBS-GFP-T into Chlora
 +
</p>
 +
 +
<p>
 +
    pTet-RBS-GFP-T into Chlora
 +
</p>
 +
 +
<p>
 +
    pTet-LacI into Chlora
 +
</p>
 +
 +
]
 +
</div>
 +
<div id="page100" class='notebook_page'>
 +
<p>
 +
    Digestion/Extraction
 +
</p>
 +
 +
<p>
 +
    GFP
 +
</p>
 +
 +
<p>
 +
    pLac-LacO
 +
</p>
 +
 +
<p dir="ltr">
 +
    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP
 +
</p>
 +
<p>
 +
    pLac-RBS-GFP-T
 +
</p>
 +
 +
<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>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    September 8
 +
</p>
 +
 +
<p>
 +
    Colonies Picked
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        14-7-5
 +
    </li>
 +
    <li>
 +
        14-7-7
 +
    </li>
 +
    <li>
 +
        14-8-2
 +
    </li>
 +
</ul>
 +
 +
</div>
 +
<div id="page102" class='notebook_page'>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    September 9, 2012
 +
</p>
 +
 +
<p>
 +
    Colony Picking
 +
</p>
 +
 +
<p>
 +
    pLac-TetR, 1 colony picked
 +
</p>
 +
 +
<p>
 +
    pLac-RBS-GFP-T, 1 colony picked
 +
</p>
 +
 +
<p>
 +
    pTet-LacI, 1 colony picked
 +
</p>
 +
 +
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<p>
 +
    pLac-LacO-GFP
 +
</p>
 +
 +
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    September 9
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock and Plasmid Isolation
 +
</p>
 +
 +
<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>
 +
    Digestion and Gel Extraction
 +
</p>
 +
 +
<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>
 +
    Ligations
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    September 10, 2012
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock/Miniprep
 +
</p>
 +
 +
<p>
 +
    pLac-TetR
 +
</p>
 +
 +
<p>
 +
    pLac-RBS-GFP-T
 +
</p>
 +
<br
 +
</div>
 +
<div id="page104" class='notebook_page'>
 +
<p>
 +
    pTet-LacI
 +
</p>
 +
 +
 +
<p>
 +
    Gel Extraction
 +
</p>
 +
 +
<p>
 +
    pLac-LacO-GFP
 +
</p>
 +
 +
<p>
 +
    pTet-LacI
 +
</p>
 +
 +
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<p>
 +
    pTet-LacI into pLac-LacO-RBS-GFP-ST
 +
</p>
 +
 +
<p>
 +
    pLac-RBS-GFP-T-pTet-LacI into chlora backbone
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate Projects
 +
</p>
 +
 +
<p>
 +
    September 10
 +
</p>
 +
 +
<p>
 +
    Colonies Picked
 +
</p>
 +
 +
<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>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        pLac
 +
    </li>
 +
    <li>
 +
        LuxR
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    September 11, 2012
 +
</p>
 +
 +
<p>
 +
    PCR
 +
</p>
 +
 +
<p>
 +
    pLac-LacO
 +
</p>
 +
 +
<p>
 +
    pLac-LacO x2
 +
</p>
 +
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    September 12, 2012
 +
</p>
 +
 +
<p>
 +
    Digestion/Gel Extraction
 +
</p>
 +
 +
<p>
 +
    pTet-LacI-pLac-LacO-RBS-GFP-ST
 +
</p>
 +
 +
 +
</div>
 +
<div id="page106" class='notebook_page'>
 +
<p>
 +
    Experiments
 +
</p>
 +
 +
<p>
 +
    Toggle with 20 micro liters IPTG
 +
</p>
 +
 +
<p>
 +
    Toggle with 40 micro liters ATc
 +
</p>
 +
 +
<p>
 +
    Strain 1 of dual population toggle with inducer strain
 +
</p>
 +
 +
<p>
 +
    Strain 2 of dual population toggle with inducer strain
 +
</p>
 +
 +
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<p>
 +
    pLac-LacO-RBS-GFP-ST-pTet-LacI into chlora backbone
 +
</p>
 +
 +
 +
<p>
 +
    Triple Population and AND Gate
 +
</p>
 +
 +
<p>
 +
    September 13
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock and Plasmid Isolation
 +
</p>
 +
 +
<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>
 +
    Innoculation
 +
</p>
 +
 +
<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>
 +
    Ligation
 +
</p>
 +
 +
<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>
 +
    Ligations for Chlora
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        I4-14-7
 +
    </li>
 +
    <li>
 +
        I4-14-8
 +
    </li>
 +
    <li>
 +
        I4-14-10
 +
    </li>
 +
</ul>
 +
<p>
 +
    Transformation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        I4-18-1
 +
    </li>
 +
    <li>
 +
        I4-18-2
 +
    </li>
 +
    <li>
 +
        I4-18-3
 +
    </li>
 +
    <li>
 +
        I401804
 +
    </li>
 +
</ul>
 +
<p>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        I3-28-2
 +
    </li>
 +
</div>
 +
<div id="page109" class='notebook_page'>
 +
    <li>
 +
        I3-30-4
 +
    </li>
 +
    <li>
 +
        I3-11-1
 +
    </li>
 +
    <li>
 +
        I3-46-4
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    September 14, 2012
 +
</p>
 +
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<p dir="ltr">
 +
    pTet-LacI-RBS-GFP(LVA)-T-T-pLac-TetR-RBS-RFP into Chlora backbone
 +
</p>
 +
<p>
 +
    pLac-RBS-GFP-T into Chlora backbone
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Glycerol Stock/Plasmid Isolation
 +
</p>
 +
 +
<p>
 +
    pLac-LacO-GFP
 +
</p>
 +
 +
 +
<p>
 +
    Digestion
 +
</p>
 +
 +
<p>
 +
    pLac-LacO-GFP
 +
</p>
 +
 +
<p>
 +
    pTet-LacI
 +
</p>
 +
 +
<p dir="ltr">
 +
    pLac-LacO x2-RBS-GFP-ST-pTet-LacI
 +
</p>
 +
 +
<p>
 +
    Inoculation
 +
</p>
 +
 +
<p>
 +
    pLac-GFP-pTet-LacI
 +
</p>
 +
 +
 +
</div>
 +
<div id="page111" class='notebook_page'>
 +
<p>
 +
    Triple Population and AND Gate
 +
</p>
 +
 +
<p>
 +
    September 15
 +
</p>
 +
 +
<p>
 +
    Plasmid Isolation
 +
</p>
 +
 +
<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>
 +
 +
<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>
 +
 +
<ul>
 +
    <li>
 +
        I4-16-4
 +
    </li>
 +
</div>
 +
<div id="page112" class='notebook_page'>
 +
    <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>
 +
 +
<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>
 +
    Innoculation
 +
</p>
 +
 +
<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>
 +
 +
 +
 +
</div>
 +
<div id="page114" class='notebook_page'>
 +
<p>
 +
    Triple Population and AND Gate
 +
</p>
 +
 +
<p>
 +
    September 18
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock and Plasmid Isolation
 +
</p>
 +
 +
<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>
 +
    Digestion and Gel Extraction
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        I4-10-7
 +
    </li>
 +
    <li>
 +
        I4-25-14
 +
    </li>
 +
    <li>
 +
        I4-25-16
 +
    </li>
 +
    <li>
 +
        I4-25-18
 +
    </li>
 +
</ul>
 +
<p>
 +
    Ligation
 +
</p>
 +
 +
<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>
 +
    Innoculation
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Pop3 AND Gate
 +
    </li>
 +
    <li>
 +
        Pop2 AND Gate
 +
    </li>
 +
    <li>
 +
        Pop1 AND Gate
 +
    </li>
 +
</ul>
 +
<p>
 +
    Testing AND Gate
 +
</p>
 +
 +
<ul>
 +
    <li>
 +
        Pop2 and 3
 +
    </li>
 +
    <li>
 +
        Pop1 and 3
 +
    </li>
 +
    <li>
 +
        Pop 1 2 3
 +
    </li>
 +
</ul>
 +
 +
<p>
 +
    Triple Population and AND Gate
 +
</p>
 +
 +
<p>
 +
    September 19
 +
</p>
 +
 +
<p>
 +
    Plate Experiments for AND Gate
 +
</p>
 +
 +
 +
 +
 +
 +
 +
</div>
 +
<div id="page116" class='notebook_page'>
 +
<p>
 +
    Triple Population and AND Gate
 +
</p>
 +
 +
<p>
 +
    September 20
 +
</p>
 +
 +
<p>
 +
    Glycerol Stock and Plasmid Isolation
 +
</p>
 +
 +
<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>
 +
 +
<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>
 +
 +
</div>
 +
<div id="page117" class='notebook_page'>
 +
<p>
 +
    Triple Population and AND Gate
 +
</p>
 +
 +
<p>
 +
    September 22
 +
</p>
 +
 +
<p>
 +
    Ligations (Repeated)
 +
</p>
 +
 +
<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>
 +
 +
<p>
 +
    Single and Double Population Projects
 +
</p>
 +
 +
<p>
 +
    September 22, 2012
 +
</p>
 +
 +
<p>
 +
    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>
 +
 +
</div>
 +
<div id="page118" class='notebook_page'>
 +
<p>
 +
    Triple Population and AND Gate
 +
</p>
 +
 +
<p>
 +
    September 22
 +
</p>
 +
 +
<p>
 +
    Ligations (Repeated)
 +
</p>
 +
 +
<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>
 +
 +
</div>
 +
<div id="page119" class='notebook_page'>
 +
<p>
 +
    Triple Population and AND Gate
 +
</p>
 +
 +
<p>
 +
    September 23
 +
</p>
 +
 +
<p>
 +
    Digestions
 +
</p>
 +
 +
<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>
 +
 +
<ul>
 +
    <li>
 +
        Pop1
 +
    </li>
 +
    <li>
 +
        Pop2
 +
    </li>
 +
    <li>
 +
        Pop3
 +
    </li>
 +
</ul>
 +
<p>
 +
    Testing AND Gate
 +
</p>
 +
 +
<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>
 +
 +
<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>
 +
 +
</div>
 +
<div id="page121" class='notebook_page'>
<p>
<p>
-
Digestion &amp; Gel Extraction
+
    Triple Population and AND Gate
</p>
</p>
 +
<p>
<p>
-
&#9679; LuxS (wrong enzymes used)
+
    September 23
</p>
</p>
 +
<p>
<p>
-
&#9679; LuxR: Concentration: 11.1 ng/&#956;l
+
    Plasmid Isolation
</p>
</p>
 +
 +
<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>
<p>
-
&#9679; LuxI: Concentration: 7.1 ng/&#956;l
+
    Digestions and Gel Extraction
</p>
</p>
-
div>
+
 
-
<div id="page3" class='notebook_page'>He determined to drop his litigation with the monastry, and relinguish his claims to the wood-cuting and fishery rihgts at once. He was the more ready to do this becuase the rights had becom much less valuable, and he had indeed the vaguest idea where the wood and river in quedtion were.
+
<ul>
-
<br><br>
+
    <li>
-
These excellant intentions were strengthed when he enterd the Father Superior's diniing-room, though, stricttly speakin, it was not a dining-room, for the Father Superior had only two rooms alltogether; they were, however, much larger and more comfortable than Father Zossima's. But tehre was was no great luxury about the furnishng of these rooms eithar. The furniture was of mohogany, covered with leather, in the old-fashionned style of 1820 the floor was not even stained, but evreything was shining with cleanlyness, and there were many chioce flowers in the windows; the most sumptuous thing in the room at the moment was, of course, the beatifuly decorated table. The cloth was clean, the service shone; there were three kinds of well-baked bread, two bottles of wine, two of excellent mead, and a large glass jug of kvas -- both the latter made in the monastery, and famous in the neigborhood. There was no vodka. Rakitin related afterwards that there were five dishes: fish-suop made of sterlets, served with little fish paties; then boiled fish served in a spesial way; then salmon cutlets, ice pudding and compote, and finally, blanc-mange. Rakitin found out about all these good things, for he could not resist peeping into the kitchen, where he already had a footing. </div>
+
        TT-pLsrA-LuxI-&gt;&gt;araC
-
<div id="page4" class='notebook_page'>He had a footting everywhere, and got informaiton about everything. He was of an uneasy and envious temper. He was well aware of his own considerable abilities, and nervously exaggerated them in his self-conceit. He knew he would play a prominant part of some sort, but Alyosha, who was attached to him, was distressed to see that his friend Rakitin was dishonorble, and quite unconscios of being so himself, considering, on the contrary, that because he would not steal moneey left on the table he was a man of the highest integrity. Neither Alyosha nor anyone else could have infleunced him in that.
+
    </li>
-
<br><br>
+
    <li>
-
Rakitin, of course, was a person of tooo little consecuense to be invited to the dinner, to which Father Iosif, Father Paissy, and one othr monk were the only inmates of the monastery invited. They were alraedy waiting when Miusov, Kalganov, and Ivan arrived. The other guest, Maximov, stood a little aside, waiting also. The Father Superior stepped into the middle of the room to receive his guests. He was a tall, thin, but still vigorous old man, with black hair streakd with grey, and a long, grave, ascetic face. He bowed to his guests in silence. But this time they approaced to receive his blessing. Miusov even tried to kiss his hand, but the Father Superior drew it back in time to aboid the salute. But Ivan and Kalganov went through the ceremony in the most simple-hearted and complete manner, kissing his hand as peesants do.</div>
+
        TT-pTetR-TetR
-
<div id="page5" class='notebook_page'>"We must apologize most humbly, your reverance," began Miusov, simpering affably, and speakin in a dignified and respecful tone. "Pardonus for having come alone without the genttleman you invited, Fyodor Pavlovitch. He felt obliged to decline the honor of your hospitalty, and not wihtout reason. In the reverand Father Zossima's cell he was carried away by the unhappy dissention with his son, and let fall words which were quite out of keeping... in fact, quite unseamly... as" -- he glanced at the monks -- "your reverance is, no doubt, already aware. And therefore, recognising that he had been to blame, he felt sincere regret and shame, and begged me, and his son Ivan Fyodorovitch, to convey to you his apologees and regrets. In brief, he hopes and desires to make amends later. He asks your blessinq, and begs you to forget what has takn place."
+
    </li>
-
<br><br>
+
    <li>
-
As he utterred the last word of his terade, Miusov completely recovered his self-complecency, and all traces of his former iritation disappaered. He fuly and sincerelly loved humanity again.</div>
+
        CP-LsrK-&gt;&gt;LuxR
-
<div id="page6" class='notebook_page'>The Father Superior listened to him with diginity, and, with a slight bend of the head, replied:
+
    </li>
-
<br><br>
+
    <li>
-
"I sincerly deplore his absence. Perhaps at our table he might have learnt to like us, and we him. Pray be seated, gentlemen."
+
        TT-pLuxR-LacI
-
<br><br>
+
    </li>
-
He stood before the holly image, and began to say grace, aloud. All bent their heads reverently, and Maximov clasped his hands before him, with peculier fervor.
+
</ul>
-
<br><br>
+
-
It was at this moment that Fyodor Pavlovitch played his last prank. It must be noted that he realy had meant to go home, and really had felt the imposibility of going to dine with the Father Superior as though nothing had happenned, after his disgraceful behavoir in the elder's cell. Not that he was so very much ashamed of himself -- quite the contrary perhaps. But still he felt it would be unseemly to go to dinner. Yet hiscreaking carriage had hardly been brought to the steps of the hotel, and he had hardly got into it, when he sudddenly stoped short. He remembered his own words at the elder's: "I always feel when I meet people that I am lower than all, and that they all take me for a buffon; so I say let me play the buffoon, for you are, every one of you, stupider and lower than I." He longed to revenge himself on everone for his own unseemliness. He suddenly recalled how he had once in the past been asked, "Why do you hate so and so, so much?" And he had answered them, with his shaemless impudence, "I'll tell you. He has done me no harm.
+
</div>
</div>
-
<div id="page7" class='notebook_page'>But I played him a dirty trick, and ever since I have hated him."
 
-
<br><br>
 
-
Rememebering that now, he smiled quietly and malignently, hesitating for a moment. His eyes gleamed, and his lips positively quivered.
 
-
<br><br>
 
-
"Well, since I have begun, I may as well go on," he decided. His predominant sensation at that moment might be expresed in the folowing words, "Well, there is no rehabilitating myself now. So let me shame them for all I am worht. I will show them I don't care what they think -- that's all!"</div>
 
-
<div id="page8" class='notebook_page'>page8 contents</div>
 
-
<div id="page9" class='notebook_page'>page9 contents</div>
 
-
<div id="page10" class='notebook_page'>page10 contents</div>
 
</html>
</html>

Latest revision as of 01:28, 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/?l

    IPTG stock: 50 ng/?l

    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/?l Tet
    • 200 ng/?l Tet
    • 300 ng/?l 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"