Team:UT-Tokyo/Notebook

From 2012.igem.org

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</p><img id="abstimg" src="https://static.igem.org/mediawiki/2012/8/8a/UT-Tokyo_LN-02.jpg" alt="box-background image" /><p>
</p><img id="abstimg" src="https://static.igem.org/mediawiki/2012/8/8a/UT-Tokyo_LN-02.jpg" alt="box-background image" /><p>
<!-- ここから</p>の前までを編集してください -->
<!-- ここから</p>の前までを編集してください -->
-
このページの概要を、簡単に記述。
 
</p></div><p></html>
</p></div><p></html>
<!-- 以下テンプレ部分まで自由記述 -->
<!-- 以下テンプレ部分まで自由記述 -->
<!-- *** What falls between these lines is the Alert Box!  You can remove it from your pages once you have read and understood the alert *** -->
<!-- *** What falls between these lines is the Alert Box!  You can remove it from your pages once you have read and understood the alert *** -->
 +
 +
== Correspondence table ==
 +
 +
{| border="1"
 +
!No.
 +
!Biobrick Parts
 +
|-
 +
|#1
 +
|pConst
 +
|-
 +
|#2
 +
|Argbox
 +
|-
 +
|#3
 +
|RBS
 +
|-
 +
|#4
 +
|Argbox×8
 +
|-
 +
|#5
 +
|d.term
 +
|-
 +
|#6
 +
|fake of fhlA
 +
|-
 +
|#7
 +
|fhlA(P-siteless)
 +
|-
 +
|#8
 +
|hycAp
 +
|-
 +
|#9
 +
|Ptrc-RBS-rocF
 +
|-
 +
|#10
 +
|fhlA(E363G)
 +
|-
 +
|#11
 +
|RFP(psb1K3)
 +
|-
 +
|#20
 +
|pLac
 +
|-
 +
|#40
 +
|RBS(weak)-GFP-d.term
 +
|-
 +
|#44
 +
|RBS(strong)-GFP-d.term
 +
|-
 +
|#66
 +
|RFP(psb1C3)
 +
|}
 +
 +
== Lab Notes ==
<html></p>
<html></p>
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<div class="LabNote">
<div class="LabNote">
-
<h2>May</h2>
+
 
 +
 
 +
<h2 id="May" name="May">May</h2>
<!--  a onclick="ShowNote('Week01');" onkeypress="ShowNote('Week01');">1st week: 5/27-6/2</a -->
<!--  a onclick="ShowNote('Week01');" onkeypress="ShowNote('Week01');">1st week: 5/27-6/2</a -->
<h3 onclick="ShowNote('Week01');" onkeypress="ShowNote('Week01');">1st week: 5/27-6/2</h3>
<h3 onclick="ShowNote('Week01');" onkeypress="ShowNote('Week01');">1st week: 5/27-6/2</h3>
<div id="Week01" class="Weeks">
<div id="Week01" class="Weeks">
<p></html>
<p></html>
 +
 +
==== May 29th ====
==== May 29th ====
*Making LB medium
*Making LB medium
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-
<h2>June</h2>
+
<h2 id="June" name="June">June</h2>
<h3 onclick="ShowNote('Week02');" onkeypress="ShowNote('Week02');">2nd week: 6/3-6/9</h3>
<h3 onclick="ShowNote('Week02');" onkeypress="ShowNote('Week02');">2nd week: 6/3-6/9</h3>
<div id="Week02" class="Weeks">
<div id="Week02" class="Weeks">
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<div id="Week03" class="Weeks">
<div id="Week03" class="Weeks">
<p></html>
<p></html>
 +
 +
none
<html></p>
<html></p>
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<h3 onclick="ShowNote('Week04');" onkeypress="ShowNote('Week04');">4th week: 6/17-6/23</h3>
<h3 onclick="ShowNote('Week04');" onkeypress="ShowNote('Week04');">4th week: 6/17-6/23</h3>
<div id="Week04" class="Weeks">
<div id="Week04" class="Weeks">
-
 
+
<p>none</p>
</div>
</div>
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-
<h2>2012/07</h2>
+
<h2 id="July" name="July">July</h2>
<h3 onclick="ShowNote('Week06');" onkeypress="ShowNote('Week06');">6th week: 7/1-7/7</h3>
<h3 onclick="ShowNote('Week06');" onkeypress="ShowNote('Week06');">6th week: 7/1-7/7</h3>
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*Miniprep
*Miniprep
** #3, #5, #20
** #3, #5, #20
 +
 +
==== July 13th ====
 +
*Nanodrop (#6).
 +
*Digestion (#3:E-P, #5:E-P, #20:E-P)
 +
*Electrophoresis (#3, #5, #20)
 +
 +
==== July 14th ====
 +
*Miniprep (#1,3,5,11,20)
 +
*Gel extraction (#6)
<html></p>
<html></p>
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<div id="Week08" class="Weeks">
<div id="Week08" class="Weeks">
<p></html>
<p></html>
 +
 +
==== July 17th ====
 +
*Transformation (#1A2,#3,#5,#20)
 +
*PCR (parts of 1. and #3,#20(Colony PCR))
 +
*Incubate #5.
 +
 +
==== July 18th ====
 +
*Miniprep (#1A2,#3,#5,#20)
 +
*PCR using u.p (#1A2,#3,#5,#20)
 +
*Colony PCR (#3,#20,H₂O)
 +
*Electrophoresis (#1A2,#3,#5,#20, #3,#20, H₂O)
 +
*PCR (2011’s #1,#36 and 2012’s #3,#5,#20, H₂O)
 +
 +
==== July 20th ====
 +
*Miniprep.
 +
*Electrophoresis (7/19’s PCR product(#1,#3,#5,#20,#36) )
 +
*Colony PCR (#3,#5,#20)
 +
 +
==== July 21st ====
 +
*Electrophoresis (7/20’s PCR product (#3,#5,#20))
 +
<html></p>
<html></p>
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<div id="Week09" class="Weeks">
<div id="Week09" class="Weeks">
<p></html>
<p></html>
 +
 +
==== July 24th ====
 +
*Making agar medium plate contain ampicillin
 +
*PCR (#6, #8)
 +
*Electrophoresis (#6, #8)
 +
*Transformation (#1)
 +
*Culture (#5, #20)
 +
 +
==== July 25th ====
 +
*Miniprep (#5, #20)
 +
*Purify (#6, #8)
 +
*Digestion (#3:S-P,#5:E-X,#11:E-S,#20:S-P)
 +
*Electrophoresis (#5, #11)
 +
*Ligation (#5,#11) (?)
 +
 +
==== July 26th ====
 +
*Culture (#1)
 +
*Digestion (#3:S-P, #5:E-X, #11:E-S, #20:S-P, fhlA:E-S, hycA:E-S)
 +
*Electrophoresis (#3:S-P, #5:E-X, #11:E-S, #20:S-P, fhlA:E-S, hycA:E-S)
 +
*Gel extraction (#3:S-P, #5:E-X, #11:E-S, #20:S-P, fhlA:E-S, hycA:E-S)
 +
 +
==== July 27th ====
 +
*Culture, Miniprep (#1)
 +
*Digestion (#5-A:E-X, #11-A:X-P)
 +
*Electrophoresis (#3:S-P, #20:S-P, #11:E-S)
 +
*Gel extraction (#3:S-P, #20:S-P, #11:E-S, E-S vector, hycAp:E-S)
 +
 +
==== July 28th ====
 +
*Digestion(#1,3,5,11,hycAp)
 +
*Gel extraction(#1,3,5,11,hycAp)
 +
*Cloning(fhlA)
 +
<html></p>
<html></p>
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-
</div>
+
<h3 onclick="ShowNote('Week10');" onkeypress="ShowNote('Week10');">10th week: 7/29-8/4</h3>
 +
<div id="Week10" class="Weeks">
 +
<p></html>
 +
==== July 30th ====
 +
*Digestion (#3:S-P, #11:X-P)
 +
*Electrophoresis (#3, #11)
 +
*Gel extraction (#3, #11)
 +
*Ligation (#3, #11)
 +
*Transformation (ligation product; #3-11)
 +
==== August 1st ====
 +
*Digestion (#3:S-P, #11:X-P)
 +
*Electrophoresis (#3, #11)
 +
*Gel extraction (#3, #11)
 +
*Ligation (#3, #11)
 +
*Transformation (ligation product; #3-11)
 +
*Miniprep (#3, #5, #11)
 +
==== August 2nd ====
 +
*Colony PCR (#3-11)
 +
*Culture (#3-11)
 +
*Colony pick up, culture (red colony of #11)
 +
*Digestion (#3:S-P, #5:E-X)
 +
 +
==== August 3th ====
 +
*Gel extraction (#3:S-P, #5:E-X)
 +
*Making solution of Kanamycin (100mg/mL)
 +
*Culture, miniprep (#11)
 +
*Culture (#11)
 +
 +
==== August 4th====
 +
*Plating, Miniprep (#11)
 +
 +
 +
<html></p>
 +
</div>
 +
 +
 +
<h2 id="August" name="August">August</h2>
 +
<h3 onclick="ShowNote('Week11');" onkeypress="ShowNote('Week11');">11th week: 8/5-8/11</h3>
 +
<div id="Week11" class="Weeks">
<p></html>
<p></html>
-
<html>
 
-
<div id="box" style="width: 700px; margin-left: 137px; padding: 5px; border: 3px solid #000; background-color: #fe2b33;">
 
-
<div id="template" style="text-align: center; font-weight: bold; font-size: large; color: #f6f6f6; padding: 5px;">
 
-
This is a template page. READ THESE INSTRUCTIONS.
 
-
</div>
 
-
<div id="instructions" style="text-align: center; font-weight: normal; font-size: small; color: #f6f6f6; padding: 5px;">
 
-
You are provided with this team page template with which to start the iGEM season.  You may choose to personalize it to fit your team but keep the same "look." Or you may choose to take your team wiki to a different level and design your own wiki.  You can find some examples <a href="https://2008.igem.org/Help:Template/Examples">HERE</a>.
 
-
</div>
 
-
<div id="warning" style="text-align: center; font-weight: bold; font-size: small; color: #f6f6f6; padding: 5px;">
 
-
You <strong>MUST</strong>  have all of the pages listed in the menu below with the names specified.  PLEASE keep all of your pages within your teams namespace. 
 
-
</div>
 
-
</div>
 
-
</html>
 
-
<!-- *** End of the alert box *** -->
+
==== August 6th====
 +
*Culture, Colony PCR (#3-11)
-
{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
+
==== August 7th====
-
!align="center"|[[Team:UT-Tokyo|Home]]
+
*Colony PCR, culture (#3-11)
-
!align="center"|[[Team:UT-Tokyo/Team|Team]]
+
*Miniprep, Digestion, Gel extraction (culture product; #3-11)
-
!align="center"|[https://igem.org/Team.cgi?year=2012&team_name=UT-Tokyo Official Team Profile]
+
-
!align="center"|[[Team:UT-Tokyo/Project|Project]]
+
==== August 8th====
-
!align="center"|[[Team:UT-Tokyo/Parts|Parts Submitted to the Registry]]
+
*Digestion (#5: E-X)
-
!align="center"|[[Team:UT-Tokyo/Modeling|Modeling]]
+
*Gel extraction (#5)
-
!align="center"|[[Team:UT-Tokyo/Notebook|Notebook]]
+
*Miniprep (#5)
-
!align="center"|[[Team:UT-Tokyo/Safety|Safety]]
+
*Colony PCR (culture product; #3-11)
-
!align="center"|[[Team:UT-Tokyo/Attributions|Attributions]]
+
-
|}
+
 +
==== August 9th====
 +
*Colony PCR (#3-11)
 +
*Culture, Miniprep (#11)
 +
*Digestion (#11:X-P)
 +
*Cloning (#6,#8)
 +
*Ligation
 +
*Gel extraction (ligation product; #5, #11)
 +
*Ligation, Transformation (#3, #11)
 +
==== August 10th====
 +
*Colony PCR (#3-11)
 +
*Miniprep (#3)
 +
*Digestion (#3:S-P)
 +
*Cloning, Colony PCR (#6,#8)
 +
*cut check, PCR check (#3-11)
 +
*Electrophoresis (ligation product; #3, #11, PCR product; #6)
 +
*Gel extraction (#6)
 +
 +
==== August 11th====
 +
*electrophoresis(PCR product; #3-11)
 +
*Digestion (#6:E-S)
 +
*electrophoresis (#8)
 +
 +
 +
<html></p>
 +
</div>
 +
 +
 +
<h3 onclick="ShowNote('Week12');" onkeypress="ShowNote('Week12');">12th week: 8/12-8/18</h3>
 +
<div id="Week12" class="Weeks">
 +
<p></html>
 +
 +
 +
==== August 13th====
 +
*Check, Culture (#3-11)
 +
*Transformation (test plasmid)
 +
 +
==== August 15th====
 +
*Gel extraction (cut check product; #3, #11)
 +
*Ligation, Transformation (#3-#11)
 +
*Make Vector for #6
 +
*Digestion (#3: E-S)
 +
*Gel extraction (digestion product; #3, #6(8/11)
 +
*PCR check (ligation product; #3-11)
 +
*Cloning (#6)
 +
 +
==== August 16th====
 +
*Gel extraction (#6)
 +
*cloning (#6)
 +
*PCR check (#3-11)
 +
*PCR-cleanup (#8)
 +
*culture (#3-11)
 +
 +
==== August 17th====
 +
*electrophoresis (#6)
 +
*Miniprep (#3-11)
 +
*Digestion (#3-#11: E-S, #6: E-S)
 +
*Gel extraction (#3-11)
 +
*Ligation, Transformation (#3-11-5)
 +
*Making solution of Ampicillin 
 +
 +
 +
<html></p>
 +
</div>
 +
 +
<h3 onclick="ShowNote('Week13');" onkeypress="ShowNote('Week13');">13th week: 8/19-8/25</h3>
 +
<div id="Week13" class="Weeks">
 +
<p></html>
 +
 +
 +
==== August 20th====
 +
*PCR check (#3-11-5)
 +
*Digestion (#8: E-S)
 +
*Cloning (#6)
 +
*Digeston (#8: E-S)
 +
*Gel extraction (#6)
 +
 +
==== August 21th====
 +
*Colony PCR (#1)
 +
*Digestion (#6: E-S, #3-11: E-S, #20: S-P)
 +
*Ligation (#3-11, #5)
 +
*Gel extraction (#3-11, #6)
 +
*Ligation (#6-pSB1A2)
 +
*Electrophresis (#3, #5; made previously)
 +
 +
==== August 22th====
 +
*Cloning (#6; from test plasmid)
 +
*Colony PCR (#3-11-5(colony is not red), #20, 8/21 ligation product;#6, #8)
 +
*Miniprep,Frozen stock (culture product; #1, #5, #20)
 +
*Transformation (#8, #3-11-5)
 +
*Digestion (#6:X-S)
 +
*Making solution of Ampicillin 
 +
*cloning retry (#6)
 +
 +
==== August 23th====
 +
*PCR check (ligation product; #6)
 +
*Cloning (#6)
 +
Colony PCR (#1, #6, #8, #11, #3-11-5, #3-11)
 +
==== August 24th====
 +
*Digestion (#6, E-S, #3-11, E-S)
 +
*Colony PCR #3-11
 +
*Electrophresis, Gel extraction (#6)
 +
*Vector insert (#6)
 +
*Sequencing (#8)
 +
 +
==== August 25th====
 +
*Digestion (#6: E-S)
 +
*Colony PCR (#6-Vector)
 +
*Gel extraction (#3-11)
 +
 +
 +
<html></p>
 +
</div>
 +
 +
<h3 onclick="ShowNote('Week14');" onkeypress="ShowNote('Week14');">14th week: 8/26-9/2</h3>
 +
<div id="Week14" class="Weeks">
 +
<p></html>
 +
 +
 +
==== August 26th====
 +
*Electrophresis (#6)
 +
*Master plate (#6)
 +
*Colony PCR (#3-11-5)
 +
*Preculture (#6)
 +
 +
==== August 27th====
 +
*Ligation (Miniprep product; #3-11, #5)
 +
*Digestion retry (#3-11, #5)
 +
*Miniprep (#6)
 +
*Mutagenesis (#6)
 +
*Sequencing (#6)
 +
 +
====August 28th ====
 +
*Mutagenesis(#6)
 +
*Digestion(#3-11,#5)
 +
 +
==== August 29th ====
 +
*Mutagenesis by inverse PCR(#6)
 +
*Digestion(#5)
 +
*Cut check(#6)
 +
*PCR check(#6)
 +
*Sequencing(#6)
 +
*Making agar medium plate contain Chloramphenicol
 +
 +
==== August 30th ====
 +
*Digestion(#5,#3-11,#8)
 +
*Sequencing(#6)
 +
*Mutagenesis(#6)
 +
 +
==== August 31st ====
 +
*Colony PCR(#6)
 +
*Digestion(#3-11,#5)
 +
*Ligation(#3-11+#5)
 +
*Transformation(#3-11-5,#66)
 +
*Culture(#66)
 +
 +
==== September 1st ====
 +
*Cloning(#6)
 +
*Miniprep(#66)
 +
 +
<html></p>
 +
</div>
 +
 +
<h2 id="September" name="September">September</h2>
 +
<h3 onclick="ShowNote('Week15');" onkeypress="ShowNote('Week15');">15th week: 9/3-9/9</h3>
 +
<div id="Week15" class="Weeks">
 +
<p></html>
 +
 +
 +
==== September 3rd ====
 +
*Miniprep(#3,#11)
 +
*Colony PCR(#3-11-5)
 +
*Transformation(#2)
 +
 +
==== September 4th ====
 +
*Digestion(#3,#6,#11)
 +
*Ligation(#3,#6)
 +
*Miniprep(#2)
 +
 +
==== September 5th ====
 +
*Ligation(#6+1A2)
 +
*Check PCR(#6)
 +
*Digestion(#2,8)
 +
 +
==== September 6th ====
 +
*Mutagenesis(#7)
 +
*Sequencing(#6)
 +
*Digestion(#5,#66)
 +
*Transformation(#2,#6)
 +
 +
==== September 7th ====
 +
*Colony PCR(#2,#8)
 +
*Cut check(EX,ES,XP,SPcut)
 +
*Digestion(#5,#66)
 +
*Ligation(#66+#5)
 +
*Transformation(#66-5)
 +
*Mutagenesis(#7)
 +
 +
==== September 8th ====
 +
*Sequencing(#6)
 +
*Colony PCR(#66-5)
 +
*Miniprep(#2)
 +
 +
<html></p>
 +
</div>
 +
 +
<h3 onclick="ShowNote('Week16');" onkeypress="ShowNote('Week16');">16th week: 9/10-9/16</h3>
 +
<div id="Week16" class="Weeks">
 +
<p></html>
 +
 +
 +
==== September 10th ====
 +
*Mutagenesis(#7)
 +
*Digestion(#2)
 +
*Transformation(#9,#40)
 +
 +
==== September 11th ====
 +
*Transformation(#44)
 +
*Mutagenesis(#7)
 +
*Miniprep(#6)
 +
*Colony PCR(#7)
 +
 +
==== September 12th ====
 +
*Culture(#44)
 +
*Miniprep(#44)
 +
*Mutagenesis(#7)
 +
*Transformation(#7)
 +
 +
==== September 13th ====
 +
*Culture(#1,#7,#20)
 +
*Colony PCR(#7)
 +
*Mutagenesis(#7)
 +
 +
==== September 14th ====
 +
*Inverse PCR(#10)
 +
*Digestion(#3,#7)
 +
*Ligation(#3-7)
 +
*Transformation(#3-7,#10)
 +
 +
==== September 15th ====
 +
*Digestion(#1,#7,#8,#44,#66)
 +
*Sequencing(#7,#10)
 +
*Colony PCR(#44,#3-7)
 +
 +
==== September 16th ====
 +
*Digestion(#1,#3,#7-5,#8,#10,#44,#66)
 +
*Transformation(#3-7-5,#3-10)
 +
 +
 +
<html></p>
 +
</div>
 +
 +
<h3 onclick="ShowNote('Week17');" onkeypress="ShowNote('Week17');">17th week: 9/17-9/23</h3>
 +
<div id="Week17" class="Weeks">
 +
<p></html>
 +
 +
==== September 17th ====
 +
*Miniprep(#7-5,#44)
 +
*Digetion(#3,5,10)
 +
*Transformation(#3-7-5,3-10-5)
 +
 +
==== September 18th ====
 +
*Miniprep(#7-5,#44)
 +
*Digetion(#3,#5,#10)
 +
*Transformation(#3-7-5,#3-10-5)
 +
 +
==== September 19th ====
 +
*Transformation(#4)
 +
*Miniprep(#3-7-5,#3-10-5)
 +
*Digestion(#3-7-5,#3-10-5,#20,#66)
 +
 +
==== September 20th ====
 +
*Miniprep(#4)
 +
*Digestion(#3-7-5,#3-10-5,#4,#20,#66)
 +
*Transformation(#20-3-7-5,#20-3-10-5,#4-4)
 +
 +
==== September 21st ====
 +
*Digestion(#3-7-5,#3-10-5,#4,#20,#66)
 +
*Transformation(#20-3-7-5,#20-3-10-5,#4-4,#7+1C3,#10+1C3)
 +
*Gas chromatography (preliminary experiment using WT)
 +
 +
==== September 22nd ====
 +
*Digestion(#3-7-5,#3-10-5,#4,#20,#66)
 +
*Transformation(#20-3-7-5,#20-3-10-5,#4-4)
 +
*Miniprep(#7+1C3,#10+1C3)
 +
 +
==== September 23rd ====
 +
*Digestion(#3-7-5,#3-10-5,#4,#8,#20,#66)
 +
*Transformation(#20-3-7-5,#20-3-10-5,#4-4)
 +
*Colony PCR(#20-3-7-5)
 +
 +
<html></p>
 +
</div>
 +
 +
<h3 onclick="ShowNote('Week18');" onkeypress="ShowNote('Week18');">18th week: 9/24-9/26</h3>
 +
<div id="Week18" class="Weeks">
 +
<p></html>
 +
 +
 +
==== September 24th ====
 +
*Digestion(#3-7-5,#3-10-5,#4,#8,#20,#66)
 +
*Transformation(#20-3-7-5,#20-3-10-5,#4-4)
 +
*Colony PCR(#20-3-7-5,#4+1C3)
 +
 +
==== September 25th ====
 +
*Digestion(#3-7-5,#3-10-5,#4,#20,#66)
 +
*Transformation(#20-3-7-5,#20-3-10-5,#4-4)
 +
*Colony PCR(#8-44)
 +
Gas chromatography (WT,#6,#10)
 +
 +
==== September 26th ====
 +
*Digestion(#20-3-7-5,#20-3-10-5,#4,#66,#8-44)
 +
*Transformation(#8-44-20-3-7-5-1C3,#8-44-20-3-10-5-1C3)
 +
*Culture(4-4-1C3)
 +
*Gas chromatography (preparation of analytical line,WT,#7,#10)
 +
 +
<html></p>
 +
</div>
 +
 +
</div>
 +
 +
 +
 +
<p></html>
-
You should make use of the calendar feature on the wiki and start a lab notebook.  This may be looked at by the judges to see how your work progressed throughout the summer.  It is a very useful organizational tool as well.
 
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Latest revision as of 03:52, 27 September 2012

Notebook

box-background image


Correspondence table

No. Biobrick Parts
#1 pConst
#2 Argbox
#3 RBS
#4 Argbox×8
#5 d.term
#6 fake of fhlA
#7 fhlA(P-siteless)
#8 hycAp
#9 Ptrc-RBS-rocF
#10 fhlA(E363G)
#11 RFP(psb1K3)
#20 pLac
#40 RBS(weak)-GFP-d.term
#44 RBS(strong)-GFP-d.term
#66 RFP(psb1C3)

Lab Notes

May

1st week: 5/27-6/2


May 29th

  • Making LB medium

May 30th

  • Making LB Agar
  • Making LB Amp Plates


May 31st

  • Preculture


June

2nd week: 6/3-6/9

June 5th

  • Plating JM109

June 6th

  • preculture

June 7th

  • Measuring OD by spectroscope
  • Meking electrocompetent cells
  • Electroporating of a test plasmid

June 8th

  • Caluculatiing transformation efficiency


3rd week: 6/10-6/16

none

4th week: 6/17-6/23

none

5th week: 6/24-6/30

June 26th

  • Making LB tetracycline plate

June 27th

June 28th

  • Making LB kanamycin plate and chloramphenicol plate
  • Transformation with a test plasmid

June 29th

  • Transformation with #1(constitutive promoter) and #11(RBS)

July

6th week: 7/1-7/7

July 3rd

  • Making K solution
  • Colony PCR DNA fragment assumed as fhlA (#6) for cloning

July 4th

  • Miniprep #1, #11

July 5th

  • Making 1% agarose solution
  • Electrophoresis (#6)
  • Digest (#1_EP, #11-_EP)

July 6th

  • Colony PCR (#6)

7th week: 7/8-7/14

July 9th

  • Electrophoresis (#6)

July 10th

  • Transformation
    • #3(RBS medium), #5(d.term), #20(pLac)

July 11th

  • Preculture
    • #3, #5, #20
  • Gel Extraction(#6)

July 12th

  • The continuation of Gel Extraction(#6)
  • Miniprep
    • #3, #5, #20

July 13th

  • Nanodrop (#6).
  • Digestion (#3:E-P, #5:E-P, #20:E-P)
  • Electrophoresis (#3, #5, #20)

July 14th

  • Miniprep (#1,3,5,11,20)
  • Gel extraction (#6)

8th week: 7/15-7/21

July 17th

  • Transformation (#1A2,#3,#5,#20)
  • PCR (parts of 1. and #3,#20(Colony PCR))
  • Incubate #5.

July 18th

  • Miniprep (#1A2,#3,#5,#20)
  • PCR using u.p (#1A2,#3,#5,#20)
  • Colony PCR (#3,#20,H₂O)
  • Electrophoresis (#1A2,#3,#5,#20, #3,#20, H₂O)
  • PCR (2011’s #1,#36 and 2012’s #3,#5,#20, H₂O)

July 20th

  • Miniprep.
  • Electrophoresis (7/19’s PCR product(#1,#3,#5,#20,#36) )
  • Colony PCR (#3,#5,#20)

July 21st

  • Electrophoresis (7/20’s PCR product (#3,#5,#20))


9th week: 7/22-7/28

July 24th

  • Making agar medium plate contain ampicillin
  • PCR (#6, #8)
  • Electrophoresis (#6, #8)
  • Transformation (#1)
  • Culture (#5, #20)

July 25th

  • Miniprep (#5, #20)
  • Purify (#6, #8)
  • Digestion (#3:S-P,#5:E-X,#11:E-S,#20:S-P)
  • Electrophoresis (#5, #11)
  • Ligation (#5,#11) (?)

July 26th

  • Culture (#1)
  • Digestion (#3:S-P, #5:E-X, #11:E-S, #20:S-P, fhlA:E-S, hycA:E-S)
  • Electrophoresis (#3:S-P, #5:E-X, #11:E-S, #20:S-P, fhlA:E-S, hycA:E-S)
  • Gel extraction (#3:S-P, #5:E-X, #11:E-S, #20:S-P, fhlA:E-S, hycA:E-S)

July 27th

  • Culture, Miniprep (#1)
  • Digestion (#5-A:E-X, #11-A:X-P)
  • Electrophoresis (#3:S-P, #20:S-P, #11:E-S)
  • Gel extraction (#3:S-P, #20:S-P, #11:E-S, E-S vector, hycAp:E-S)

July 28th

  • Digestion(#1,3,5,11,hycAp)
  • Gel extraction(#1,3,5,11,hycAp)
  • Cloning(fhlA)


10th week: 7/29-8/4

July 30th

  • Digestion (#3:S-P, #11:X-P)
  • Electrophoresis (#3, #11)
  • Gel extraction (#3, #11)
  • Ligation (#3, #11)
  • Transformation (ligation product; #3-11)

August 1st

  • Digestion (#3:S-P, #11:X-P)
  • Electrophoresis (#3, #11)
  • Gel extraction (#3, #11)
  • Ligation (#3, #11)
  • Transformation (ligation product; #3-11)
  • Miniprep (#3, #5, #11)

August 2nd

  • Colony PCR (#3-11)
  • Culture (#3-11)
  • Colony pick up, culture (red colony of #11)
  • Digestion (#3:S-P, #5:E-X)

August 3th

  • Gel extraction (#3:S-P, #5:E-X)
  • Making solution of Kanamycin (100mg/mL)
  • Culture, miniprep (#11)
  • Culture (#11)

August 4th

  • Plating, Miniprep (#11)


August

11th week: 8/5-8/11

August 6th

  • Culture, Colony PCR (#3-11)

August 7th

  • Colony PCR, culture (#3-11)
  • Miniprep, Digestion, Gel extraction (culture product; #3-11)

August 8th

  • Digestion (#5: E-X)
  • Gel extraction (#5)
  • Miniprep (#5)
  • Colony PCR (culture product; #3-11)

August 9th

  • Colony PCR (#3-11)
  • Culture, Miniprep (#11)
  • Digestion (#11:X-P)
  • Cloning (#6,#8)
  • Ligation
  • Gel extraction (ligation product; #5, #11)
  • Ligation, Transformation (#3, #11)

August 10th

  • Colony PCR (#3-11)
  • Miniprep (#3)
  • Digestion (#3:S-P)
  • Cloning, Colony PCR (#6,#8)
  • cut check, PCR check (#3-11)
  • Electrophoresis (ligation product; #3, #11, PCR product; #6)
  • Gel extraction (#6)

August 11th

  • electrophoresis(PCR product; #3-11)
  • Digestion (#6:E-S)
  • electrophoresis (#8)


12th week: 8/12-8/18


August 13th

  • Check, Culture (#3-11)
  • Transformation (test plasmid)

August 15th

  • Gel extraction (cut check product; #3, #11)
  • Ligation, Transformation (#3-#11)
  • Make Vector for #6
  • Digestion (#3: E-S)
  • Gel extraction (digestion product; #3, #6(8/11)
  • PCR check (ligation product; #3-11)
  • Cloning (#6)

August 16th

  • Gel extraction (#6)
  • cloning (#6)
  • PCR check (#3-11)
  • PCR-cleanup (#8)
  • culture (#3-11)

August 17th

  • electrophoresis (#6)
  • Miniprep (#3-11)
  • Digestion (#3-#11: E-S, #6: E-S)
  • Gel extraction (#3-11)
  • Ligation, Transformation (#3-11-5)
  • Making solution of Ampicillin


13th week: 8/19-8/25


August 20th

  • PCR check (#3-11-5)
  • Digestion (#8: E-S)
  • Cloning (#6)
  • Digeston (#8: E-S)
  • Gel extraction (#6)

August 21th

  • Colony PCR (#1)
  • Digestion (#6: E-S, #3-11: E-S, #20: S-P)
  • Ligation (#3-11, #5)
  • Gel extraction (#3-11, #6)
  • Ligation (#6-pSB1A2)
  • Electrophresis (#3, #5; made previously)

August 22th

  • Cloning (#6; from test plasmid)
  • Colony PCR (#3-11-5(colony is not red), #20, 8/21 ligation product;#6, #8)
  • Miniprep,Frozen stock (culture product; #1, #5, #20)
  • Transformation (#8, #3-11-5)
  • Digestion (#6:X-S)
  • Making solution of Ampicillin
  • cloning retry (#6)

August 23th

  • PCR check (ligation product; #6)
  • Cloning (#6)

Colony PCR (#1, #6, #8, #11, #3-11-5, #3-11)

August 24th

  • Digestion (#6, E-S, #3-11, E-S)
  • Colony PCR #3-11
  • Electrophresis, Gel extraction (#6)
  • Vector insert (#6)
  • Sequencing (#8)

August 25th

  • Digestion (#6: E-S)
  • Colony PCR (#6-Vector)
  • Gel extraction (#3-11)


14th week: 8/26-9/2


August 26th

  • Electrophresis (#6)
  • Master plate (#6)
  • Colony PCR (#3-11-5)
  • Preculture (#6)

August 27th

  • Ligation (Miniprep product; #3-11, #5)
  • Digestion retry (#3-11, #5)
  • Miniprep (#6)
  • Mutagenesis (#6)
  • Sequencing (#6)

August 28th

  • Mutagenesis(#6)
  • Digestion(#3-11,#5)

August 29th

  • Mutagenesis by inverse PCR(#6)
  • Digestion(#5)
  • Cut check(#6)
  • PCR check(#6)
  • Sequencing(#6)
  • Making agar medium plate contain Chloramphenicol

August 30th

  • Digestion(#5,#3-11,#8)
  • Sequencing(#6)
  • Mutagenesis(#6)

August 31st

  • Colony PCR(#6)
  • Digestion(#3-11,#5)
  • Ligation(#3-11+#5)
  • Transformation(#3-11-5,#66)
  • Culture(#66)

September 1st

  • Cloning(#6)
  • Miniprep(#66)

September

15th week: 9/3-9/9


September 3rd

  • Miniprep(#3,#11)
  • Colony PCR(#3-11-5)
  • Transformation(#2)

September 4th

  • Digestion(#3,#6,#11)
  • Ligation(#3,#6)
  • Miniprep(#2)

September 5th

  • Ligation(#6+1A2)
  • Check PCR(#6)
  • Digestion(#2,8)

September 6th

  • Mutagenesis(#7)
  • Sequencing(#6)
  • Digestion(#5,#66)
  • Transformation(#2,#6)

September 7th

  • Colony PCR(#2,#8)
  • Cut check(EX,ES,XP,SPcut)
  • Digestion(#5,#66)
  • Ligation(#66+#5)
  • Transformation(#66-5)
  • Mutagenesis(#7)

September 8th

  • Sequencing(#6)
  • Colony PCR(#66-5)
  • Miniprep(#2)

16th week: 9/10-9/16


September 10th

  • Mutagenesis(#7)
  • Digestion(#2)
  • Transformation(#9,#40)

September 11th

  • Transformation(#44)
  • Mutagenesis(#7)
  • Miniprep(#6)
  • Colony PCR(#7)

September 12th

  • Culture(#44)
  • Miniprep(#44)
  • Mutagenesis(#7)
  • Transformation(#7)

September 13th

  • Culture(#1,#7,#20)
  • Colony PCR(#7)
  • Mutagenesis(#7)

September 14th

  • Inverse PCR(#10)
  • Digestion(#3,#7)
  • Ligation(#3-7)
  • Transformation(#3-7,#10)

September 15th

  • Digestion(#1,#7,#8,#44,#66)
  • Sequencing(#7,#10)
  • Colony PCR(#44,#3-7)

September 16th

  • Digestion(#1,#3,#7-5,#8,#10,#44,#66)
  • Transformation(#3-7-5,#3-10)


17th week: 9/17-9/23

September 17th

  • Miniprep(#7-5,#44)
  • Digetion(#3,5,10)
  • Transformation(#3-7-5,3-10-5)

September 18th

  • Miniprep(#7-5,#44)
  • Digetion(#3,#5,#10)
  • Transformation(#3-7-5,#3-10-5)

September 19th

  • Transformation(#4)
  • Miniprep(#3-7-5,#3-10-5)
  • Digestion(#3-7-5,#3-10-5,#20,#66)

September 20th

  • Miniprep(#4)
  • Digestion(#3-7-5,#3-10-5,#4,#20,#66)
  • Transformation(#20-3-7-5,#20-3-10-5,#4-4)

September 21st

  • Digestion(#3-7-5,#3-10-5,#4,#20,#66)
  • Transformation(#20-3-7-5,#20-3-10-5,#4-4,#7+1C3,#10+1C3)
  • Gas chromatography (preliminary experiment using WT)

September 22nd

  • Digestion(#3-7-5,#3-10-5,#4,#20,#66)
  • Transformation(#20-3-7-5,#20-3-10-5,#4-4)
  • Miniprep(#7+1C3,#10+1C3)

September 23rd

  • Digestion(#3-7-5,#3-10-5,#4,#8,#20,#66)
  • Transformation(#20-3-7-5,#20-3-10-5,#4-4)
  • Colony PCR(#20-3-7-5)

18th week: 9/24-9/26


September 24th

  • Digestion(#3-7-5,#3-10-5,#4,#8,#20,#66)
  • Transformation(#20-3-7-5,#20-3-10-5,#4-4)
  • Colony PCR(#20-3-7-5,#4+1C3)

September 25th

  • Digestion(#3-7-5,#3-10-5,#4,#20,#66)
  • Transformation(#20-3-7-5,#20-3-10-5,#4-4)
  • Colony PCR(#8-44)

Gas chromatography (WT,#6,#10)

September 26th

  • Digestion(#20-3-7-5,#20-3-10-5,#4,#66,#8-44)
  • Transformation(#8-44-20-3-7-5-1C3,#8-44-20-3-10-5-1C3)
  • Culture(4-4-1C3)
  • Gas chromatography (preparation of analytical line,WT,#7,#10)


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