Team:ZJU-China/Notebook/August

From 2011.igem.org

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<h3>Week2</h3><hr/>
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  <tr><td width="76">Day</td><td width="349">Note</td></tr>
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    <td id="sheetleft">Jul.11th Monday</td>
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    <td><table id="intable" width="541" border="0" cellspacing="0" cellpadding="1">
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    <td width="128">•  Set up new LB culture plates with ampicillin and kanamycin</p></td>
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    <td width="196">•
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      ·Sterilization of Glycerol and <br />
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        ·Preparation of 25mg/mL kanamycin</td>
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  <td>•Transformation of the parts mentioned on Jul.9th for the second time</td>
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  <td>•Observation the sections</td>
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    <td id="sheetleft">Jul 12th Tuesday
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    <td><table id="intable" width="560" border="0" cellspacing="0" cellpadding="1">
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    <td width="128">• Pick two colonies of each parts and cultivate them in LB medium</td>
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    <td width="203"> •Transformation of three parts(20J,20H.22B from Kit plates of 2011 Distribution ) which are related to the degradation of Cellulose </td>
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  <td width="91">•·Mini  prep to isolate 10I,12I and 22M</td>
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  <td width="130">•Conservation of 10I,12I,22M and 11P</td>
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    <td id="sheetleft">Jul.13th
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Wednesday
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</td>
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    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
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    <td width="120">• Place the culture plate of 20J,20H and 22B in the fridge.</td>
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    <td width="102">•Min prep to isolate 13K
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•Conservation of 13K
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<td width="156">•Restriction digest of the parts(12I,10I,22M,13K) with EcoRI and PstI</td>
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<td width="95">•Gel electrophoresis to analyse restriction fragments</td>
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<td width="157">•Test the Tm of Primers CP1&amp;CS,NP&amp;NS with 13K. The result of Gel electrophoresis shows that 60.2℃ is the Tm of NS and NP, and 57.4℃ is the Tm of CS and CP1.</td>
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    <td id="sheetleft">Jul.14th
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Thursday
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    <td><table width="618" border="0" cellspacing="0" cellpadding="1">
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    <td >• Test the Tm of the YFP,RFP,tetR,Vgb and nirB primers.
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The result of Gel electrophoresis shows that the Tm for the primers of Vgb is 54℃ and the Tm for the primers of nirB is 55℃.The RCR of YFP,RFP and tetR failed.
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    <td id="sheetleft">Jul.15th
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Friday
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    <td><table width="600" border="0" cellspacing="0" cellpadding="1">
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        <td width="155"> • PCR(Phusion)<br/>
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• Digest 10I, 22M, 13K
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• Used wrong cutter, digestion again.
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</td>
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<td>• Run the results of PCR and the first digestion. The annealing temperature of YFP needed change. The digestion results confirmed</td>
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<td>• Run the digestion results of second time. The bands are confirmed.</td>
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    <td id="sheetleft">Jul.16th
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Saturday
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    <td width="127">• Cut the linearized pSB1k3 with E+P</td>
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    <td width="123">• Purify the digestion results of 22M, 13K, 10I</td>
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    <td width="133">• Confirm digestion of pSB1k3 by electrophoresis, then purification</td>
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    <td width="113">• Test Tm of YFP<br/>
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• ligation: 22M+10I, 13K+10I
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<td width="114">• Tm of YFP is 54 degree
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• transform the ligation results.
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    <td id="sheetleft">Jul.17th
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Sunday
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        <td>• PCR 22M<br/>
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• purify Vgb, RFP, RFP-tetR, YFP-tetR. Gibson PCR.
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</td>
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<td>• ligation the purified the fragments in yesterday.</td>
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<td>• 22M PCR<br/>
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• Transform the ligation results.
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<p>&nbsp;</p>
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Revision as of 17:36, 2 October 2011

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Labnote

 Biobrick group | Biofilm group 

This group...........

Week1

DayNote
Jul.4th Monday
• Aerobic cultivation of DH5α • Preparation of apparatus for the formation of biofilm
Jul 5th Tuesday  
Jul.6th Wednesday
•Receiving primers ordered previously
Jul.7th Thursday
• Preparation of the aliquot of the primers • Something wrong with a shaking incubator
Jul.8th Friday
• Preparation of culture plates for the transformations
Jul.9th Saturday
• Preparation of culture plates for the transformations • protocols of transformation
Jul.10th Sunday
• Several colonies were picked up and cultivated in 5mL LB medium. •Cryosectioning of biofilm

 

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