Team:UT-Tokyo/Notebook
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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 --> | ||
- | |||
</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> | + | |
- | <a onclick="ShowNote('Week01');" onkeypress="ShowNote('Week01');">1st week: 5/27-6/2</a> | + | |
+ | <h2 id="May" name="May">May</h2> | ||
+ | <!-- 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 ==== |
+ | *Making LB medium | ||
+ | |||
+ | ==== May 30th ==== | ||
+ | *Making LB Agar | ||
+ | *Making LB Amp Plates | ||
+ | |||
+ | |||
+ | ==== May 31st ==== | ||
+ | *Preculture | ||
Line 35: | Line 93: | ||
- | <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"> | ||
<p></html> | <p></html> | ||
- | ==== | + | ==== 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 | ||
Line 58: | Line 117: | ||
<h3 onclick="ShowNote('Week03');" onkeypress="ShowNote('Week03');">3rd week: 6/10-6/16</h3> | <h3 onclick="ShowNote('Week03');" onkeypress="ShowNote('Week03');">3rd week: 6/10-6/16</h3> | ||
<div id="Week03" class="Weeks"> | <div id="Week03" class="Weeks"> | ||
+ | <p></html> | ||
+ | none | ||
+ | |||
+ | <html></p> | ||
</div> | </div> | ||
<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> | ||
- | <h3 onclick="ShowNote('Week05');" onkeypress="ShowNote('Week05');"> | + | <h3 onclick="ShowNote('Week05');" onkeypress="ShowNote('Week05');">5th week: 6/24-6/30</h3> |
<div id="Week05" class="Weeks"> | <div id="Week05" class="Weeks"> | ||
+ | <p></html> | ||
+ | ==== 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) | ||
+ | |||
+ | <html></p> | ||
+ | |||
</div> | </div> | ||
- | <h2> | + | <h2 id="July" name="July">July</h2> |
- | <h3 onclick="ShowNote('Week06');" onkeypress="ShowNote(' | + | <h3 onclick="ShowNote('Week06');" onkeypress="ShowNote('Week06');">6th week: 7/1-7/7</h3> |
<div id="Week06" class="Weeks"> | <div id="Week06" class="Weeks"> | ||
+ | <p></html> | ||
+ | |||
+ | ==== 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) | ||
+ | |||
+ | <html></p> | ||
</div> | </div> | ||
- | |||
+ | <h3 onclick="ShowNote('Week07');" onkeypress="ShowNote('Week07');">7th week: 7/8-7/14</h3> | ||
+ | <div id="Week07" class="Weeks"> | ||
+ | <p></html> | ||
+ | ==== 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) | ||
+ | |||
+ | <html></p> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | <h3 onclick="ShowNote('Week08');" onkeypress="ShowNote('Week08');">8th week: 7/15-7/21</h3> | ||
+ | <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> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | <h3 onclick="ShowNote('Week09');" onkeypress="ShowNote('Week09');">9th week: 7/22-7/28</h3> | ||
+ | <div id="Week09" class="Weeks"> | ||
+ | <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> | ||
+ | </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> | ||
+ | |||
+ | ==== 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) | ||
+ | |||
+ | |||
+ | <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> | ||
- | |||
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<html></p> | <html></p> |
Latest revision as of 03:52, 27 September 2012
Notebook
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)