Team:ZJU-China/Notebook
From 2011.igem.org
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- | <body><div id="contentwrapper"> | + | <body><div id="contentwrapper"> |
- | <div class="main"> | + | <div class="main"> |
- | <table id="tab" border="0" cellspacing="1" cellpadding="1"> | + | <table id="tab" border="0" cellspacing="1" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td><div class="pagetitle" id="stra1"><a> <h3>Biobrick</h3> | + | <td><div class="pagetitle" id="stra1"><a> <h3>Biobrick</h3> |
- | <p>brief introduction</p></a></div></td> | + | <p>brief introduction</p></a></div></td> |
- | <td><div class="pagetitle" id="stra2"><a href="https://2011.igem.org/Team:ZJU-China/Notebook/Biofilm"> <h3>Biofilm Formation</h3> | + | <td><div class="pagetitle" id="stra2"><a href="https://2011.igem.org/Team:ZJU-China/Notebook/Biofilm"> <h3>Biofilm Formation</h3> |
- | <p>brief introduction</p></a></div></td> | + | <p>brief introduction</p></a></div></td> |
- | </tr> | + | </tr> |
- | </table> | + | </table> |
<div id="leftul"> | <div id="leftul"> | ||
<ul class="kwicks"> | <ul class="kwicks"> | ||
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<li id="kwick3"><a>week3</a></li> | <li id="kwick3"><a>week3</a></li> | ||
<li id="kwick4"><a>week4</a></li> | <li id="kwick4"><a>week4</a></li> | ||
- | <li id="kwick5"><a>week5</a></li> | + | <li id="kwick5"><a>week5</a></li> |
- | <li id="kwick6"><a>week6</a></li> | + | <li id="kwick6"><a>week6</a></li> |
- | <li id="kwick7"><a>week7</a></li> | + | <li id="kwick7"><a>week7</a></li> |
- | <li id="kwick8"><a>week8</a></li> | + | <li id="kwick8"><a>week8</a></li> |
- | <li id="kwick9"><a>week9</a></li> | + | <li id="kwick9"><a>week9</a></li> |
- | <li id="kwick10"><a>week10</a></li> | + | <li id="kwick10"><a>week10</a></li> |
- | <li id="kwick11"><a>week11</a></li> | + | <li id="kwick11"><a>week11</a></li> |
- | <li id="kwick12"><a>week12</a></li> | + | <li id="kwick12"><a>week12</a></li> |
- | <li id=" | + | <li id="afkwick12"><a style="font-size:18pt;margin:1px;padding:1px;">after week12</a></li> |
</ul> | </ul> | ||
</div> | </div> | ||
- | <div class="bigblock"><div class="inner" id="week1"> | + | <div class="bigblock"><div class="inner" id="week1"> |
- | + | ||
- | <div class="block" id="nsheet"> | + | <div class="block" id="nsheet"> |
- | <h3>Week1</h3><hr/> | + | <h3>Week1</h3><hr/> |
- | + | ||
- | <table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1"> | + | <table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1"> |
- | <tr><td width="76">Day</td><td width="349">Note</td></tr> | + | <tr><td width="76">Day</td><td width="349">Note</td></tr> |
- | <tr> | + | <tr> |
- | <td id="sheetleft">Jul.4th Monday</td> | + | <td id="sheetleft">Jul.4th Monday</td> |
- | <td><table id="intable" width="328" border="0" cellspacing="0" cellpadding="1"> | + | <td><table id="intable" width="328" border="0" cellspacing="0" cellpadding="1"> |
- | <td width="128">• Aerobic cultivation of DH5α</td> | + | <td width="128">• Aerobic cultivation of DH5α</td> |
- | <td width="196">• Preparation of apparatus for the formation of biofilm</td> | + | <td width="196">• Preparation of apparatus for the formation of biofilm</td> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul 5th Tuesday | <td id="sheetleft">Jul 5th Tuesday | ||
- | </td> | + | </td> |
- | <td> </td> | + | <td> </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.6th | <td id="sheetleft">Jul.6th | ||
Wednesday | Wednesday | ||
- | </td> | + | </td> |
- | <td><table width="217" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="217" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td width="215">•Receiving primers ordered previously</td> | + | <td width="215">•Receiving primers ordered previously</td> |
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.7th | <td id="sheetleft">Jul.7th | ||
Thursday | Thursday | ||
- | </td> | + | </td> |
- | <td><table width="238" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="238" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td width="200">• Preparation of the aliquot of the primers</td> | + | <td width="200">• Preparation of the aliquot of the primers</td> |
- | <td width="200">• Something wrong with a shaking incubator</td> | + | <td width="200">• Something wrong with a shaking incubator</td> |
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.8th | <td id="sheetleft">Jul.8th | ||
Friday | Friday | ||
- | </td> | + | </td> |
- | <td><table width="222" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="222" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td width="155">• Preparation of culture plates for the transformations</td> | + | <td width="155">• Preparation of culture plates for the transformations</td> |
- | </tr> | + | </tr> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.9th | <td id="sheetleft">Jul.9th | ||
Saturday | Saturday | ||
- | </td> | + | </td> |
- | <td><table width="313" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="313" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td width="127">• Preparation of culture plates for the transformations</td> | + | <td width="127">• Preparation of culture plates for the transformations</td> |
- | <td width="182">• protocols of transformation</td> | + | <td width="182">• protocols of transformation</td> |
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.10th | <td id="sheetleft">Jul.10th | ||
Sunday | Sunday | ||
- | </td> | + | </td> |
- | <td><table width="246" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="246" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
<td>• Several colonies were picked up and cultivated in 5mL LB medium. | <td>• Several colonies were picked up and cultivated in 5mL LB medium. | ||
•Cryosectioning of biofilm | •Cryosectioning of biofilm | ||
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | + | ||
- | </table> | + | </table> |
- | <p> </p> | + | <p> </p> |
- | </div> | + | </div> |
- | + | ||
- | <div class="block" id="summ"> | + | <div class="block" id="summ"> |
- | <h3 style="color:white;">Data&protocol</h3> | + | <h3 style="color:white;">Data&protocol</h3> |
- | <hr/> | + | <hr/> |
- | <div class="block" id="monday"> | + | <div class="block" id="monday"> |
- | <h3>Monday</h3> | + | <h3>Monday</h3> |
- | <hr/> | + | <hr/> |
- | <img src="https://static.igem.org/mediawiki/2011/6/61/IMG_8382.jpg" style="margin-left:100px;" alt="blanknote" /> | + | <img src="https://static.igem.org/mediawiki/2011/6/61/IMG_8382.jpg" style="margin-left:100px;" alt="blanknote" /> |
- | <p>There is no data in first week</p> | + | <p>There is no data in first week</p> |
- | </div> | + | </div> |
- | <p> </p> | + | <p> </p> |
- | </div> | + | </div> |
- | </div> | + | </div> |
- | <!-- 11111111111111111111111111111111111111111111111111 --> | + | <!-- 11111111111111111111111111111111111111111111111111 --> |
- | <div class="inner" id="week2"> | + | <div class="inner" id="week2"> |
- | <div class="block" id="nsheet"> | + | <div class="block" id="nsheet"> |
- | <h3>Week2</h3><hr/> | + | <h3>Week2</h3><hr/> |
- | <table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1"> | + | <table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1"> |
- | <tr><td width="76">Day</td><td width="349">Note</td></tr> | + | <tr><td width="76">Day</td><td width="349">Note</td></tr> |
- | <tr> | + | <tr> |
- | <td id="sheetleft">Jul.11th Monday</td> | + | <td id="sheetleft">Jul.11th Monday</td> |
- | <td><table id="intable" width="541" border="0" cellspacing="0" cellpadding="1"> | + | <td><table id="intable" width="541" border="0" cellspacing="0" cellpadding="1"> |
- | <td width="128">• Set up new LB culture plates with ampicillin and kanamycin</p></td> | + | <td width="128">• Set up new LB culture plates with ampicillin and kanamycin</p></td> |
<td width="196">• | <td width="196">• | ||
- | ·Sterilization of Glycerol and <br /> | + | ·Sterilization of Glycerol and <br /> |
- | ·Preparation of 25mg/mL kanamycin</td> | + | ·Preparation of 25mg/mL kanamycin</td> |
- | <td>•Transformation of the parts mentioned on Jul.9th for the second time</td> | + | <td>•Transformation of the parts mentioned on Jul.9th for the second time</td> |
- | <td>•Observation the sections</td> | + | <td>•Observation the sections</td> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul 12th Tuesday | <td id="sheetleft">Jul 12th Tuesday | ||
- | </td> | + | </td> |
- | <td><table id="intable" width="560" border="0" cellspacing="0" cellpadding="1"> | + | <td><table id="intable" width="560" border="0" cellspacing="0" cellpadding="1"> |
- | <td width="128">• Pick two colonies of each parts and cultivate them in LB medium</td> | + | <td width="128">• Pick two colonies of each parts and cultivate them in LB medium</td> |
- | <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> | + | <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> |
- | <td width="91">•·Mini prep to isolate 10I,12I and 22M</td> | + | <td width="91">•·Mini prep to isolate 10I,12I and 22M</td> |
- | <td width="130">•Conservation of 10I,12I,22M and 11P</td> | + | <td width="130">•Conservation of 10I,12I,22M and 11P</td> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.13th | <td id="sheetleft">Jul.13th | ||
Wednesday | Wednesday | ||
- | </td> | + | </td> |
- | <td><table width="640" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="640" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td width="120">• Place the culture plate of 20J,20H and 22B in the fridge.</td> | + | <td width="120">• Place the culture plate of 20J,20H and 22B in the fridge.</td> |
<td width="102">•Min prep to isolate 13K | <td width="102">•Min prep to isolate 13K | ||
•Conservation of 13K | •Conservation of 13K | ||
- | </td> | + | </td> |
- | <td width="156">•Restriction digest of the parts(12I,10I,22M,13K) with EcoRI and PstI</td> | + | <td width="156">•Restriction digest of the parts(12I,10I,22M,13K) with EcoRI and PstI</td> |
- | <td width="95">•Gel electrophoresis to analyse restriction fragments</td> | + | <td width="95">•Gel electrophoresis to analyse restriction fragments</td> |
- | <td width="157">•Test the Tm of Primers CP1&CS,NP&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> | + | <td width="157">•Test the Tm of Primers CP1&CS,NP&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> |
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.14th | <td id="sheetleft">Jul.14th | ||
Thursday | Thursday | ||
- | </td> | + | </td> |
- | <td><table width="618" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="618" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
<td >• Test the Tm of the YFP,RFP,tetR,Vgb and nirB primers. | <td >• Test the Tm of the YFP,RFP,tetR,Vgb and nirB primers. | ||
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. | 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. | ||
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.15th | <td id="sheetleft">Jul.15th | ||
Friday | Friday | ||
- | </td> | + | </td> |
- | <td><table width="600" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="600" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td width="155"> • PCR(Phusion)<br/> | + | <td width="155"> • PCR(Phusion)<br/> |
• Digest 10I, 22M, 13K | • Digest 10I, 22M, 13K | ||
• Used wrong cutter, digestion again. | • Used wrong cutter, digestion again. | ||
- | </td> | + | </td> |
- | <td>• Run the results of PCR and the first digestion. The annealing temperature of YFP needed change. The digestion results confirmed</td> | + | <td>• Run the results of PCR and the first digestion. The annealing temperature of YFP needed change. The digestion results confirmed</td> |
- | <td>• Run the digestion results of second time. The bands are confirmed.</td> | + | <td>• Run the digestion results of second time. The bands are confirmed.</td> |
- | </tr> | + | </tr> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.16th | <td id="sheetleft">Jul.16th | ||
Saturday | Saturday | ||
- | </td> | + | </td> |
- | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td width="127">• Cut the linearized pSB1k3 with E+P</td> | + | <td width="127">• Cut the linearized pSB1k3 with E+P</td> |
- | <td width="123">• Purify the digestion results of 22M, 13K, 10I</td> | + | <td width="123">• Purify the digestion results of 22M, 13K, 10I</td> |
- | <td width="133">• Confirm digestion of pSB1k3 by electrophoresis, then purification</td> | + | <td width="133">• Confirm digestion of pSB1k3 by electrophoresis, then purification</td> |
- | <td width="113">• Test Tm of YFP<br/> | + | <td width="113">• Test Tm of YFP<br/> |
• ligation: 22M+10I, 13K+10I | • ligation: 22M+10I, 13K+10I | ||
- | </td> | + | </td> |
<td width="114">• Tm of YFP is 54 degree | <td width="114">• Tm of YFP is 54 degree | ||
• transform the ligation results. | • transform the ligation results. | ||
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.17th | <td id="sheetleft">Jul.17th | ||
Sunday | Sunday | ||
- | </td> | + | </td> |
- | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td>• PCR 22M<br/> | + | <td>• PCR 22M<br/> |
• purify Vgb, RFP, RFP-tetR, YFP-tetR. Gibson PCR. | • purify Vgb, RFP, RFP-tetR, YFP-tetR. Gibson PCR. | ||
- | </td> | + | </td> |
- | <td>• ligation the purified the fragments in yesterday.</td> | + | <td>• ligation the purified the fragments in yesterday.</td> |
- | <td>• 22M PCR<br/> | + | <td>• 22M PCR<br/> |
• Transform the ligation results. | • Transform the ligation results. | ||
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | + | ||
- | </table> | + | </table> |
- | <p> </p> | + | <p> </p> |
- | + | ||
- | + | ||
- | </div> | + | </div> |
- | <div class="block" id="summ"> | + | <div class="block" id="summ"> |
- | <h3 style="color:white;">Data&protocol</h3> | + | <h3 style="color:white;">Data&protocol</h3> |
- | <hr/> | + | <hr/> |
- | <div class="block" id="monday"> | + | <div class="block" id="monday"> |
- | <h3>Jul.13th Wednesday</h3> | + | <h3>Jul.13th Wednesday</h3> |
- | <hr/> | + | <hr/> |
- | <p><strong>Systems of restriction digestion with EcoRI and PstI</strong></p> | + | <p><strong>Systems of restriction digestion with EcoRI and PstI</strong></p> |
- | + | ||
- | <table style="margin:15px;" width="110" border="1" cellspacing="0" cellpadding="1"> | + | <table style="margin:15px;" width="110" border="1" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td>Plasmid</td> | + | <td>Plasmid</td> |
- | <td>1μL (>100ng)</td> | + | <td>1μL (>100ng)</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>EcoRI</td> | + | <td>EcoRI</td> |
- | <td>1μL</td> | + | <td>1μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>PstI</td> | + | <td>PstI</td> |
- | <td>1μL</td> | + | <td>1μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>10×Buffer Tango</td> | + | <td>10×Buffer Tango</td> |
- | <td>2μL</td> | + | <td>2μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>ddH2O</td> | + | <td>ddH2O</td> |
- | <td>15μL</td> | + | <td>15μL</td> |
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | <img src="https://static.igem.org/mediawiki/2011/c/c3/0713program.png" /> | + | <img src="https://static.igem.org/mediawiki/2011/c/c3/0713program.png" /> |
- | + | ||
- | <p><strong>Temperature grad</strong><br/>56℃,57.4℃,60.2℃,62.9℃,64.3℃,65.8℃</p> | + | <p><strong>Temperature grad</strong><br/>56℃,57.4℃,60.2℃,62.9℃,64.3℃,65.8℃</p> |
- | + | ||
- | <p> </p> | + | <p> </p> |
- | + | ||
- | <p><strong>PCR system (test the Tm of the primers CP1&CS, NP&NS)</strong></p> | + | <p><strong>PCR system (test the Tm of the primers CP1&CS, NP&NS)</strong></p> |
- | + | ||
- | <table style="margin:15px;" width="110" border="1" cellspacing="0" cellpadding="1"> | + | <table style="margin:15px;" width="110" border="1" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td>10×Buffer</td> | + | <td>10×Buffer</td> |
- | <td> 2μL</td> | + | <td> 2μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>dNTPs</td> | + | <td>dNTPs</td> |
- | <td>0.5μL</td> | + | <td>0.5μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>primers CP1 (NP)</td> | + | <td>primers CP1 (NP)</td> |
- | <td>1μL</td> | + | <td>1μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>primers CS (NS)</td> | + | <td>primers CS (NS)</td> |
- | <td>1μL</td> | + | <td>1μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>Template </td> | + | <td>Template </td> |
- | <td>1μL</td> | + | <td>1μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>rTaq </td> | + | <td>rTaq </td> |
- | <td>0.2μL</td> | + | <td>0.2μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>H2O </td> | + | <td>H2O </td> |
- | <td>14.5μL</td> | + | <td>14.5μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>Total</td> | + | <td>Total</td> |
- | <td> 20μL</td> | + | <td> 20μL</td> |
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | + | ||
- | </div> | + | </div> |
- | <div class="block" id="tuesday"> | + | <div class="block" id="tuesday"> |
- | <h3>Jul.14th Thursday</h3> | + | <h3>Jul.14th Thursday</h3> |
- | <hr/> | + | <hr/> |
- | <p>PCR systems(test the Tm of the primers for YFP,RFP,tetR,Vgb and nirB )</p> | + | <p>PCR systems(test the Tm of the primers for YFP,RFP,tetR,Vgb and nirB )</p> |
- | <table width="110" border="1" cellspacing="0" cellpadding="1"> | + | <table width="110" border="1" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td>10×Buffer</td> | + | <td>10×Buffer</td> |
- | <td> 2μL</td> | + | <td> 2μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>dNTPs</td> | + | <td>dNTPs</td> |
- | <td>0.5μL</td> | + | <td>0.5μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>primers F </td> | + | <td>primers F </td> |
- | <td>0.5μL</td> | + | <td>0.5μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>primers R </td> | + | <td>primers R </td> |
- | <td>0.5μL</td> | + | <td>0.5μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>Template </td> | + | <td>Template </td> |
- | <td>1μL</td> | + | <td>1μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>Taq </td> | + | <td>Taq </td> |
- | <td>0.2μL</td> | + | <td>0.2μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>H2O </td> | + | <td>H2O </td> |
- | <td>15.5μL</td> | + | <td>15.5μL</td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
- | <td>Total</td> | + | <td>Total</td> |
- | <td> 20μL</td> | + | <td> 20μL</td> |
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | <p> </p> | + | <p> </p> |
- | <img src="http://ung.igem.org/wiki/images/2/20/0714program.png" /> | + | <img src="http://ung.igem.org/wiki/images/2/20/0714program.png" /> |
- | <p>Temperature grad and the results:</p> | + | <p>Temperature grad and the results:</p> |
- | <img src="https://static.igem.org/mediawiki/2011/6/6b/0714tempera.png" /> | + | <img src="https://static.igem.org/mediawiki/2011/6/6b/0714tempera.png" /> |
- | <p>vgb&nirB pcr test:</p> | + | <p>vgb&nirB pcr test:</p> |
- | <img src="https://static.igem.org/mediawiki/2011/7/74/20110714_vgb%26nirB_pcr_test.jpg" /> | + | <img src="https://static.igem.org/mediawiki/2011/7/74/20110714_vgb%26nirB_pcr_test.jpg" /> |
- | <p> </p> | + | <p> </p> |
- | </div> | + | </div> |
- | <p> </p> | + | <p> </p> |
- | </div> | + | </div> |
- | </div> | + | </div> |
- | <!-- 2222222222222222222222222222222222222222222 --> | + | <!-- 2222222222222222222222222222222222222222222 --> |
- | <div class="inner" id="week3"> | + | <div class="inner" id="week3"> |
- | <div class="block" id="nsheet"> | + | <div class="block" id="nsheet"> |
- | <h3>Week3</h3><hr/> | + | <h3>Week3</h3><hr/> |
- | + | ||
- | <table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1"> | + | <table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1"> |
- | <tr><td width="76">Day</td><td width="349">Note</td></tr> | + | <tr><td width="76">Day</td><td width="349">Note</td></tr> |
- | <tr> | + | <tr> |
- | <td id="sheetleft">Jul.18th Monday</td> | + | <td id="sheetleft">Jul.18th Monday</td> |
- | <td><table id="intable" width="541" border="0" cellspacing="0" cellpadding="1"> | + | <td><table id="intable" width="541" border="0" cellspacing="0" cellpadding="1"> |
- | <td width="128">• Genome extraction of E.coli </td> | + | <td width="128">• Genome extraction of E.coli </td> |
- | <td width="196">• PCR of NirB<br /> | + | <td width="196">• PCR of NirB<br /> |
- | • cut 10I with X+P, cut 22M, 13K with E+S, cut pSB1k3 with E+P</td> | + | • cut 10I with X+P, cut 22M, 13K with E+S, cut pSB1k3 with E+P</td> |
- | <td>• Ligate: 10I+13K, 10I+22M <br/> | + | <td>• Ligate: 10I+13K, 10I+22M <br/> |
• Repeat NirB PCR | • Repeat NirB PCR | ||
- | </td> | + | </td> |
- | <td><p align="left">• Culture 11P<br /> | + | <td><p align="left">• Culture 11P<br /> |
- | • Miniprep 10I, 22M, 13K</p></td> | + | • Miniprep 10I, 22M, 13K</p></td> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul 19th Tuesday | <td id="sheetleft">Jul 19th Tuesday | ||
- | </td> | + | </td> |
- | <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1"> | + | <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1"> |
- | <td width="140">• mini-prep: 10I+13K, 10I+22M</td> | + | <td width="140">• mini-prep: 10I+13K, 10I+22M</td> |
- | <td width="213"> •insert 10I+13K, 10I+22M into pSB1k3 </td> | + | <td width="213"> •insert 10I+13K, 10I+22M into pSB1k3 </td> |
- | <td width="110">•transform: 5E, 3C, 7C, 1K, 1I from the distribution plate</td> | + | <td width="110">•transform: 5E, 3C, 7C, 1K, 1I from the distribution plate</td> |
- | <td width="144">•transform 10I+22M, 10I+13K</td> | + | <td width="144">•transform 10I+22M, 10I+13K</td> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.20th | <td id="sheetleft">Jul.20th | ||
Wednesday | Wednesday | ||
- | </td> | + | </td> |
- | <td><table width="640" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="640" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td width="170">•Colonies of 22M+10I, 13K+10I, 5E, 3C, 7C, 1K, 1I confirmed</td> | + | <td width="170">•Colonies of 22M+10I, 13K+10I, 5E, 3C, 7C, 1K, 1I confirmed</td> |
<td width="102">• Gibson PCR | <td width="102">• Gibson PCR | ||
- | </td> | + | </td> |
- | <td >•Colony PCR 22M+10I, 13K+10I, 5E, 3C, 7C, 1K, 1I</td> | + | <td >•Colony PCR 22M+10I, 13K+10I, 5E, 3C, 7C, 1K, 1I</td> |
- | <td width="95">•PCR NirB</td> | + | <td width="95">•PCR NirB</td> |
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.21th | <td id="sheetleft">Jul.21th | ||
Thursday | Thursday | ||
- | </td> | + | </td> |
- | <td><table width="618" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="618" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
<td >•PCR verification of 22M+10I, 13K+10I, 5E, 3C, 7C, 1K, 1I | <td >•PCR verification of 22M+10I, 13K+10I, 5E, 3C, 7C, 1K, 1I | ||
- | </td> | + | </td> |
<td>• Gibson PCR | <td>• Gibson PCR | ||
• Run the results of PCR verification. Bands confirmed. | • Run the results of PCR verification. Bands confirmed. | ||
- | </td> | + | </td> |
<td>• Culture the 22M+10I, 13K+10I, 5E, 3C, 7C, 1K, 1I | <td>• Culture the 22M+10I, 13K+10I, 5E, 3C, 7C, 1K, 1I | ||
• Purification of Gibson PCR results | • Purification of Gibson PCR results | ||
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.22th | <td id="sheetleft">Jul.22th | ||
Friday | Friday | ||
- | </td> | + | </td> |
- | <td><table width="627" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="627" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
<td width="117"> • PCR amplification of Gibson assembly results | <td width="117"> • PCR amplification of Gibson assembly results | ||
- | </td> | + | </td> |
- | <td width="144">• Mini prep: 22M+10I, 22.Presever in -20<br/> | + | <td width="144">• Mini prep: 22M+10I, 22.Presever in -20<br/> |
- | • Medi prep: 1K,1I,3C,5E,7C<br/> | + | • Medi prep: 1K,1I,3C,5E,7C<br/> |
• Gibson Assembly fail. | • Gibson Assembly fail. | ||
- | </td> | + | </td> |
- | <td width="164">• PCR NirB by Phusion<br/> | + | <td width="164">• PCR NirB by Phusion<br/> |
- | •Repeat PCR by changing Pnibr to Gnirbr<br/> | + | •Repeat PCR by changing Pnibr to Gnirbr<br/> |
•Cut the mini and medi prep results with E | •Cut the mini and medi prep results with E | ||
- | </td> | + | </td> |
- | <td width="194">•Run the results of PCR and digestion. Fail in PCR of NirB, succeed in 10I+3K and 10I+22M ligation.</td> | + | <td width="194">•Run the results of PCR and digestion. Fail in PCR of NirB, succeed in 10I+3K and 10I+22M ligation.</td> |
- | </tr> | + | </tr> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.23th | <td id="sheetleft">Jul.23th | ||
Saturday | Saturday | ||
- | </td> | + | </td> |
- | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
<td >•Double digestion of 13K+10I, 22M+10I, pSB1c3 | <td >•Double digestion of 13K+10I, 22M+10I, pSB1c3 | ||
- | + | ||
- | </td> | + | </td> |
- | <td >• Fail in purification of Medi prep</td> | + | <td >• Fail in purification of Medi prep</td> |
- | <td >• PCR NirB<br/> | + | <td >• PCR NirB<br/> |
• Ligate NirB+13K+10I, Vgb+22M+10I | • Ligate NirB+13K+10I, Vgb+22M+10I | ||
- | </td> | + | </td> |
- | + | ||
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.24th | <td id="sheetleft">Jul.24th | ||
Sunday | Sunday | ||
- | </td> | + | </td> |
- | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td width="244">• Gibson assembly: NirB+RFP+tetR<br/> | + | <td width="244">• Gibson assembly: NirB+RFP+tetR<br/> |
• Cut results of Gibson assembly and pSB1c3. | • Cut results of Gibson assembly and pSB1c3. | ||
- | + | ||
- | </td> | + | </td> |
- | <td width="164">• Purify the ligation results in yesterday</td> | + | <td width="164">• Purify the ligation results in yesterday</td> |
<td width="206">• ligate with backbone | <td width="206">• ligate with backbone | ||
• Culture 1I, 1K, 3C, 5E, 7C, 11P | • Culture 1I, 1K, 3C, 5E, 7C, 11P | ||
- | + | ||
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | + | ||
- | </table> | + | </table> |
- | <p> </p> | + | <p> </p> |
- | </div> | + | </div> |
- | <div class="block" id="summ"> | + | <div class="block" id="summ"> |
- | <h3 style="color:white;">Data&protocol</h3> | + | <h3 style="color:white;">Data&protocol</h3> |
- | <hr/> | + | <hr/> |
- | <div class="block" id="monday"> | + | <div class="block" id="monday"> |
- | <h3>Monday</h3> | + | <h3>Monday</h3> |
- | <hr/> | + | <hr/> |
- | <p>WEEK3</p> | + | <p>WEEK3</p> |
- | </div> | + | </div> |
- | <div class="block" id="tuesday"> | + | <div class="block" id="tuesday"> |
- | <h3>Tuesday</h3> | + | <h3>Tuesday</h3> |
- | <hr/> | + | <hr/> |
- | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> | + | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> |
- | <p> </p> | + | <p> </p> |
- | </div> | + | </div> |
- | </div> | + | </div> |
- | <!-- 3333333333333333333333333333333333333 --> | + | <!-- 3333333333333333333333333333333333333 --> |
- | <div class="inner" id="week4"> | + | <div class="inner" id="week4"> |
- | <div class="block" id="nsheet"> | + | <div class="block" id="nsheet"> |
- | <h3>Week4</h3><hr/> | + | <h3>Week4</h3><hr/> |
- | + | ||
- | <table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1"> | + | <table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1"> |
- | <tr><td width="76">Day</td><td width="349">Note</td></tr> | + | <tr><td width="76">Day</td><td width="349">Note</td></tr> |
- | <tr> | + | <tr> |
- | <td id="sheetleft">Jul.25th Monday</td> | + | <td id="sheetleft">Jul.25th Monday</td> |
- | <td><table id="intable" width="541" border="0" cellspacing="0" cellpadding="1"> | + | <td><table id="intable" width="541" border="0" cellspacing="0" cellpadding="1"> |
- | <td width="128">• Transform Pnirb+13k+10I+pSBK3-2</td> | + | <td width="128">• Transform Pnirb+13k+10I+pSBK3-2</td> |
- | <td >• Place the culture in 4 degree</td> | + | <td >• Place the culture in 4 degree</td> |
<td>• Medi-and mini-prep of five cultre | <td>• Medi-and mini-prep of five cultre | ||
- | </td> | + | </td> |
- | <td>•Gibson Assembly confirmation by gel</td> | + | <td>•Gibson Assembly confirmation by gel</td> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul. 26th Tuesday | <td id="sheetleft">Jul. 26th Tuesday | ||
- | </td> | + | </td> |
- | <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1"> | + | <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1"> |
- | <td width="140">• Cut Pnirb, Pvgb,(E+S)<br/> | + | <td width="140">• Cut Pnirb, Pvgb,(E+S)<br/> |
• Cut 10I+13K, 10I+22M.(X+P) | • Cut 10I+13K, 10I+22M.(X+P) | ||
- | </td> | + | </td> |
- | <td width="181" > • Ligation: vgb+10I+22M, nirB+10I+13K,</td> | + | <td width="181" > • Ligation: vgb+10I+22M, nirB+10I+13K,</td> |
- | <td width="142">• Ligation: pSBK13+vgb+10I+22M, pSBK13+nirB+10I+13K</td> | + | <td width="142">• Ligation: pSBK13+vgb+10I+22M, pSBK13+nirB+10I+13K</td> |
- | <td width="144">• Transform the ligation results.</td> | + | <td width="144">• Transform the ligation results.</td> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.27th | <td id="sheetleft">Jul.27th | ||
Wednesday | Wednesday | ||
- | </td> | + | </td> |
- | <td><table width="640" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="640" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td width="170">• No colony on the plate<br/> | + | <td width="170">• No colony on the plate<br/> |
• Gibson assembly | • Gibson assembly | ||
- | </td> | + | </td> |
<td width="184">• Run the results of Gibson assembly | <td width="184">• Run the results of Gibson assembly | ||
- | </td> | + | </td> |
- | <td width="280" >• Excise the bands, the purify the DNA<br/> | + | <td width="280" >• Excise the bands, the purify the DNA<br/> |
• Gibson Assenmly | • Gibson Assenmly | ||
- | </td> | + | </td> |
- | + | ||
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.28th | <td id="sheetleft">Jul.28th | ||
Thursday | Thursday | ||
- | </td> | + | </td> |
- | <td><table width="618" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="618" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
<td >•2011 China Meet-up @ Hefei | <td >•2011 China Meet-up @ Hefei | ||
- | </td> | + | </td> |
- | + | ||
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.29th | <td id="sheetleft">Jul.29th | ||
Friday | Friday | ||
- | </td> | + | </td> |
- | <td><table width="627" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="627" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td >•2011 China Meet-up @ Hefei</td> | + | <td >•2011 China Meet-up @ Hefei</td> |
- | </tr> | + | </tr> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.30th | <td id="sheetleft">Jul.30th | ||
Saturday | Saturday | ||
- | </td> | + | </td> |
- | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | + | ||
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Jul.31th | <td id="sheetleft">Jul.31th | ||
Sunday | Sunday | ||
- | </td> | + | </td> |
- | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
<td width="244">• Culture 22M+10I, 13K+10I | <td width="244">• Culture 22M+10I, 13K+10I | ||
- | + | ||
- | </td> | + | </td> |
- | <td width="164">• Cut a0H, 20J, 22B</td> | + | <td width="164">• Cut a0H, 20J, 22B</td> |
- | + | ||
- | </tr> | + | </tr> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | + | ||
- | </table> | + | </table> |
- | <p> </p> | + | <p> </p> |
- | </div> | + | </div> |
- | + | ||
- | <div class="block" id="summ"> | + | <div class="block" id="summ"> |
- | + | ||
- | <h3 style="color:white;">Data&protocol</h3> | + | <h3 style="color:white;">Data&protocol</h3> |
- | <hr/> | + | <hr/> |
- | <div class="block" id="monday"> | + | <div class="block" id="monday"> |
- | <h3>Monday</h3> | + | <h3>Monday</h3> |
- | <hr/> | + | <hr/> |
- | <p>WEEK3</p> | + | <p>WEEK3</p> |
- | </div> | + | </div> |
- | <div class="block" id="tuesday"> | + | <div class="block" id="tuesday"> |
- | <h3>Tuesday</h3> | + | <h3>Tuesday</h3> |
- | <hr/> | + | <hr/> |
- | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> | + | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> |
- | <p> </p> | + | <p> </p> |
- | </div> | + | </div> |
- | </div> | + | </div> |
- | <!-- 444444444444444444444444444444444 --> | + | <!-- 444444444444444444444444444444444 --> |
- | <div class="inner" id="week5"> | + | <div class="inner" id="week5"> |
- | + | ||
- | <div class="block" id="nsheet"> | + | <div class="block" id="nsheet"> |
- | <h3>Week5</h3><hr/> | + | <h3>Week5</h3><hr/> |
- | + | ||
- | <table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1"> | + | <table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1"> |
- | <tr><td width="76">Day</td><td width="349">Note</td></tr> | + | <tr><td width="76">Day</td><td width="349">Note</td></tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Aug.1st | <td id="sheetleft">Aug.1st | ||
Monday | Monday | ||
- | </td> | + | </td> |
- | <td><table id="intable" width="541" border="0" cellspacing="0" cellpadding="1"> | + | <td><table id="intable" width="541" border="0" cellspacing="0" cellpadding="1"> |
<td width="286">•Fusion PCR | <td width="286">•Fusion PCR | ||
Vgb+YFP+tetR, NirB+RFP+tetR, | Vgb+YFP+tetR, NirB+RFP+tetR, | ||
- | </td> | + | </td> |
- | <td width="251" >•PCR: NirB, Vgb</td> | + | <td width="251" >•PCR: NirB, Vgb</td> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Aug.2nd | <td id="sheetleft">Aug.2nd | ||
Tuesday | Tuesday | ||
- | + | ||
- | </td> | + | </td> |
- | <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1"> | + | <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1"> |
<td width="140"> •Cut: 13K+10I, 22M+10I | <td width="140"> •Cut: 13K+10I, 22M+10I | ||
- | </td> | + | </td> |
- | <td width="181" > • Purify: 13K+10I, 22M+10I<br/> | + | <td width="181" > • Purify: 13K+10I, 22M+10I<br/> |
• Ligation: Pvgb+22M+10I, Pnirb+13K+10I | • Ligation: Pvgb+22M+10I, Pnirb+13K+10I | ||
- | </td> | + | </td> |
- | <td width="142">• PCR backbones<br/> | + | <td width="142">• PCR backbones<br/> |
• Electrophresis | • Electrophresis | ||
- | </td> | + | </td> |
- | <td width="144">• Gel excision and purification</td> | + | <td width="144">• Gel excision and purification</td> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Aug.3rd | <td id="sheetleft">Aug.3rd | ||
Wednesday | Wednesday | ||
- | + | ||
- | </td> | + | </td> |
- | <td><table width="640" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="640" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
<td width="170">• Make Phusion Buffer, 5×isothermel buffer | <td width="170">• Make Phusion Buffer, 5×isothermel buffer | ||
- | </td> | + | </td> |
<td width="184">• colony PCR: 20H, 20J, 22B | <td width="184">• colony PCR: 20H, 20J, 22B | ||
- | </td> | + | </td> |
- | + | ||
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Aug.4th | <td id="sheetleft">Aug.4th | ||
Thursday | Thursday | ||
- | + | ||
- | </td> | + | </td> |
- | <td><table width="618" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="618" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
<td >• Cut: 10I+22M, 10I+13; and purification | <td >• Cut: 10I+22M, 10I+13; and purification | ||
- | </td> | + | </td> |
- | <td>• Ligation: Pvgb+22M+10I, Pnirb+13K+10I,<br/> | + | <td>• Ligation: Pvgb+22M+10I, Pnirb+13K+10I,<br/> |
• colony PCR: 13K+10I | • colony PCR: 13K+10I | ||
- | </td> | + | </td> |
- | <td>• Culture: 20H, 20J, 22B, 1K, 1I,3C, 5E, 7C</td> | + | <td>• Culture: 20H, 20J, 22B, 1K, 1I,3C, 5E, 7C</td> |
- | + | ||
- | <td>• Transform: 1G, 3A, 5A, 7A from the distribution plate</td> | + | <td>• Transform: 1G, 3A, 5A, 7A from the distribution plate</td> |
- | + | ||
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Aug.5th | <td id="sheetleft">Aug.5th | ||
Friday | Friday | ||
- | + | ||
- | </td> | + | </td> |
- | <td><table width="627" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="627" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td >• Check the plates. Contamination, or no positive colonies</td> | + | <td >• Check the plates. Contamination, or no positive colonies</td> |
- | <td>• Miniprep: 5 backbones, 22B-1, 22B-3</td> | + | <td>• Miniprep: 5 backbones, 22B-1, 22B-3</td> |
- | <td>• PCR: nirB from 13K+10I+nirB to firm the ligation. One positive result.</td> | + | <td>• PCR: nirB from 13K+10I+nirB to firm the ligation. One positive result.</td> |
- | <td>• Culture the positive colony.</td> | + | <td>• Culture the positive colony.</td> |
- | </tr> | + | </tr> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Aug.6th | <td id="sheetleft">Aug.6th | ||
Saturday | Saturday | ||
- | + | ||
- | </td> | + | </td> |
- | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | + | ||
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Aug.7th | <td id="sheetleft">Aug.7th | ||
Sunday | Sunday | ||
- | </td> | + | </td> |
- | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td width="244">• Cut: 22M,13K with E & S | + | <td width="244">• Cut: 22M,13K with E & S |
- | + | ||
- | </td> | + | </td> |
- | <td width="164">• Culture the red colonies from plate of pSB1C3</td> | + | <td width="164">• Culture the red colonies from plate of pSB1C3</td> |
- | <td>• PCR: 5 backbones<br/> | + | <td>• PCR: 5 backbones<br/> |
• Cut the PCR products with P+E and run the gel to confirm. | • Cut the PCR products with P+E and run the gel to confirm. | ||
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | + | ||
- | </table> | + | </table> |
- | + | ||
- | </div> | + | </div> |
- | + | ||
- | <div class="block" id="summ"> | + | <div class="block" id="summ"> |
- | <h3 style="color:white;">Data&protocol</h3> | + | <h3 style="color:white;">Data&protocol</h3> |
- | <hr/> | + | <hr/> |
- | <div class="block" id="monday"> | + | <div class="block" id="monday"> |
- | <h3>Monday</h3> | + | <h3>Monday</h3> |
- | <hr/> | + | <hr/> |
- | <p>WEEK3</p> | + | <p>WEEK3</p> |
- | </div> | + | </div> |
- | <div class="block" id="tuesday"> | + | <div class="block" id="tuesday"> |
- | <h3>Tuesday</h3> | + | <h3>Tuesday</h3> |
- | <hr/> | + | <hr/> |
- | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> | + | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> |
- | <p> </p> | + | <p> </p> |
- | </div> | + | </div> |
- | </div> | + | </div> |
- | <!-- 55555555555555555555555555555555555555 --> | + | <!-- 55555555555555555555555555555555555555 --> |
- | <div class="inner" id="week6"> | + | <div class="inner" id="week6"> |
- | <div class="block" id="nsheet"> | + | <div class="block" id="nsheet"> |
- | <h3>Week6</h3><hr/> | + | <h3>Week6</h3><hr/> |
- | + | ||
- | <table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1"> | + | <table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1"> |
- | <tr><td width="76">Day</td><td width="349">Note</td></tr> | + | <tr><td width="76">Day</td><td width="349">Note</td></tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Aut.8th | <td id="sheetleft">Aut.8th | ||
Monday | Monday | ||
- | + | ||
- | </td> | + | </td> |
- | <td><table id="intable" width="603" border="0" cellspacing="0" cellpadding="1"> | + | <td><table id="intable" width="603" border="0" cellspacing="0" cellpadding="1"> |
<td width="191">• The gel results of 5 backbones are right. | <td width="191">• The gel results of 5 backbones are right. | ||
- | </td> | + | </td> |
- | <td width="249" >• Miniprep: pSB1C3, vgb+22M+10I</td><br /> | + | <td width="249" >• Miniprep: pSB1C3, vgb+22M+10I</td><br /> |
- | <td width="157">• Culture vgb+22M+1oI in hypoxia<br/> | + | <td width="157">• Culture vgb+22M+1oI in hypoxia<br/> |
•PCR pSB1C3 | •PCR pSB1C3 | ||
- | </td> | + | </td> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Aug.9th | <td id="sheetleft">Aug.9th | ||
Tuesday | Tuesday | ||
- | + | ||
- | + | ||
- | </td> | + | </td> |
- | <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1"> | + | <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1"> |
<td width="140"> • Run the PCR product | <td width="140"> • Run the PCR product | ||
- | </td> | + | </td> |
<td width="181" > • PCR pSB1C3 again | <td width="181" > • PCR pSB1C3 again | ||
- | </td> | + | </td> |
<td width="142">• Run the product, results are good. | <td width="142">• Run the product, results are good. | ||
- | </td> | + | </td> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Aug.10th | <td id="sheetleft">Aug.10th | ||
Wednesday | Wednesday | ||
- | + | ||
- | + | ||
- | </td> | + | </td> |
- | <td><table width="640" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="640" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td width="170">• Culture: 20H, 20J<br/> | + | <td width="170">• Culture: 20H, 20J<br/> |
• Transform 13K from plate 3 | • Transform 13K from plate 3 | ||
- | + | ||
- | </td> | + | </td> |
<td width="184">• Miniprep: 20H-1, 20J-1 | <td width="184">• Miniprep: 20H-1, 20J-1 | ||
- | </td> | + | </td> |
- | <td>• Cut: the 20H-1, 20J-1, 22B-1, 22B-3 with E+P</td> | + | <td>• Cut: the 20H-1, 20J-1, 22B-1, 22B-3 with E+P</td> |
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Aug.11th | <td id="sheetleft">Aug.11th | ||
Thursday | Thursday | ||
- | + | ||
- | + | ||
- | </td> | + | </td> |
- | <td><table width="618" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="618" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
<td >• Run the digestion results. Bands are confirmed right. | <td >• Run the digestion results. Bands are confirmed right. | ||
- | </td> | + | </td> |
<td>• Colony PCR vgb+22M+10I | <td>• Colony PCR vgb+22M+10I | ||
- | </td> | + | </td> |
<td>• Cut the plasmid of vgb+22M+10I | <td>• Cut the plasmid of vgb+22M+10I | ||
- | + | ||
- | </td> | + | </td> |
- | + | ||
- | + | ||
- | </tr> | + | </tr> |
- | </table> | + | </table> |
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Aug.12th | <td id="sheetleft">Aug.12th | ||
Friday | Friday | ||
- | + | ||
- | + | ||
- | </td> | + | </td> |
- | <td><table width="627" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="627" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
- | <td >• Make new stock of competent cells</td> | + | <td >• Make new stock of competent cells</td> |
- | <td>• Ligation: 13K+10I<br/> | + | <td>• Ligation: 13K+10I<br/> |
• Cut the PCR results and 22M with E+P | • Cut the PCR results and 22M with E+P | ||
- | </td> | + | </td> |
- | <td>• Sequence: nirB, vgbL, 12I, 20H, 20J, 22B</td> | + | <td>• Sequence: nirB, vgbL, 12I, 20H, 20J, 22B</td> |
- | <td>• Transform 13K+10I+pSB1K3<br/> | + | <td>• Transform 13K+10I+pSB1K3<br/> |
• Cut 13K to validate. Results are right. | • Cut 13K to validate. Results are right. | ||
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | <tr> | + | <tr> |
<td id="sheetleft">Aug.13th | <td id="sheetleft">Aug.13th | ||
- | |||
- | + | Saturday | |
- | </td> | + | |
- | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> | + | |
- | <tr> | + | </td> |
- | <td>• Colony PCR: 13K+10I+pSB1K3</td> | + | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> |
- | <td>• Culture: vgb+22M+10I in 5ml(shaking), 5ml(water bath), 10ml(water bath), 15(water bath).</td> | + | <tr> |
- | <td>• Check the YFP expression of 5ml(shaking) and 15ml(water bath). No yellow fluorescent cells,</td> | + | <td>• Colony PCR: 13K+10I+pSB1K3</td> |
- | + | <td>• Culture: vgb+22M+10I in 5ml(shaking), 5ml(water bath), 10ml(water bath), 15(water bath).</td> | |
- | </tr> | + | <td>• Check the YFP expression of 5ml(shaking) and 15ml(water bath). No yellow fluorescent cells,</td> |
- | </table> | + | |
- | </td> | + | </tr> |
- | </tr> | + | </table> |
- | <tr> | + | </td> |
+ | </tr> | ||
+ | <tr> | ||
<td id="sheetleft">Aug.14th | <td id="sheetleft">Aug.14th | ||
Sunday | Sunday | ||
- | + | ||
- | </td> | + | </td> |
- | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> | + | <td><table width="620" border="0" cellspacing="0" cellpadding="1"> |
- | <tr> | + | <tr> |
<td width="244">• Transform: the Gibson assembly results. | <td width="244">• Transform: the Gibson assembly results. | ||
- | + | ||
- | </td> | + | </td> |
- | <td width="164">• Miniprep: 13K+10I<br/> | + | <td width="164">• Miniprep: 13K+10I<br/> |
• Cut: 13K+10I | • Cut: 13K+10I | ||
- | </td> | + | </td> |
- | <td>• Purification: the digestion results.<br/> | + | <td>• Purification: the digestion results.<br/> |
• Ligation: nirB+13K+10I | • Ligation: nirB+13K+10I | ||
- | + | ||
- | </td> | + | </td> |
- | </tr> | + | </tr> |
- | </table></td> | + | </table></td> |
- | </tr> | + | </tr> |
- | + | ||
- | </table> | + | </table> |
- | + | ||
- | </div> | + | </div> |
- | + | ||
- | <div class="block" id="summ"> | + | <div class="block" id="summ"> |
- | <h3 style="color:white;">Data&protocol</h3> | + | <h3 style="color:white;">Data&protocol</h3> |
- | <hr/> | + | <hr/> |
- | <div class="block" id="monday"> | + | <div class="block" id="monday"> |
- | <h3>Monday</h3> | + | <h3>Monday</h3> |
- | <hr/> | + | <hr/> |
- | <p>WEEK3</p> | + | <p>WEEK3</p> |
- | </div> | + | </div> |
- | <div class="block" id="tuesday"> | + | <div class="block" id="tuesday"> |
- | <h3>Tuesday</h3> | + | <h3>Tuesday</h3> |
- | <hr/> | + | <hr/> |
- | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> | + | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> |
- | <p> </p> | + | <p> </p> |
- | </div> | + | </div> |
- | </div> | + | </div> |
- | <!-- 666666666666666666666666666666666 --> | + | <!-- 666666666666666666666666666666666 --> |
- | <div class="inner" id="week7"> | + | <div class="inner" id="week7"> |
- | <div class="block" id="summ"> | + | <div class="block" id="summ"> |
- | <h3 style="color:white;">Data&protocol</h3> | + | <h3 style="color:white;">Data&protocol</h3> |
- | <hr/> | + | <hr/> |
- | <div class="block" id="monday"> | + | <div class="block" id="monday"> |
- | <h3>Monday</h3> | + | <h3>Monday</h3> |
- | <hr/> | + | <hr/> |
- | <p> | + | <p>WEEK7</p> |
- | </div> | + | </div> |
- | <div class="block" id="tuesday"> | + | <div class="block" id="tuesday"> |
- | <h3>Tuesday</h3> | + | <h3>Tuesday</h3> |
- | <hr/> | + | <hr/> |
- | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> | + | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> |
- | <p> </p> | + | <p> </p> |
- | </div> | + | </div> |
- | </div> | + | </div> |
- | <!-- 7777777777777777777777777777 --> | + | <!-- 7777777777777777777777777777 --> |
- | <div class="inner" id="week8"> | + | <div class="inner" id="week8"> |
- | <div class="block" id="summ"> | + | <div class="block" id="summ"> |
- | <h3 style="color:white;">Data&protocol</h3> | + | <h3 style="color:white;">Data&protocol</h3> |
- | <hr/> | + | <hr/> |
- | <div class="block" id="monday"> | + | <div class="block" id="monday"> |
- | <h3>Monday</h3> | + | <h3>Monday</h3> |
- | <hr/> | + | <hr/> |
- | <p> | + | <p>WEEK8</p> |
- | </div> | + | </div> |
- | <div class="block" id="tuesday"> | + | <div class="block" id="tuesday"> |
- | <h3>Tuesday</h3> | + | <h3>Tuesday</h3> |
- | <hr/> | + | <hr/> |
- | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> | + | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> |
- | <p> </p> | + | <p> </p> |
- | </div> | + | </div> |
- | </div> | + | </div> |
- | <!-- 8888888888888888888888888888 --> | + | <!-- 8888888888888888888888888888 --> |
- | <div class="inner" id="week9"> | + | <div class="inner" id="week9"> |
- | <div class="block" id="summ"> | + | <div class="block" id="summ"> |
- | <h3 style="color:white;">Data&protocol</h3> | + | <h3 style="color:white;">Data&protocol</h3> |
- | <hr/> | + | <hr/> |
- | <div class="block" id="monday"> | + | <div class="block" id="monday"> |
- | <h3>Monday</h3> | + | <h3>Monday</h3> |
- | <hr/> | + | <hr/> |
- | <p> | + | <p>WEEK9</p> |
- | </div> | + | </div> |
- | <div class="block" id="tuesday"> | + | <div class="block" id="tuesday"> |
- | <h3>Tuesday</h3> | + | <h3>Tuesday</h3> |
- | <hr/> | + | <hr/> |
- | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> | + | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> |
- | <p> </p> | + | <p> </p> |
- | </div> | + | </div> |
- | </div> | + | </div> |
- | <!-- 9999999999999999999999999999 --> | + | <!-- 9999999999999999999999999999 --> |
- | <div class="inner" id="week10"> | + | <div class="inner" id="week10"> |
- | <div class="block" id="summ"> | + | <div class="block" id="summ"> |
- | <h3 style="color:white;">Data&protocol</h3> | + | <h3 style="color:white;">Data&protocol</h3> |
- | <hr/> | + | <hr/> |
- | <div class="block" id="monday"> | + | <div class="block" id="monday"> |
- | <h3>Monday</h3> | + | <h3>Monday</h3> |
- | <hr/> | + | <hr/> |
- | <p> | + | <p>WEEK10</p> |
- | </div> | + | </div> |
- | <div class="block" id="tuesday"> | + | <div class="block" id="tuesday"> |
- | <h3>Tuesday</h3> | + | <h3>Tuesday</h3> |
- | <hr/> | + | <hr/> |
- | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> | + | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> |
- | <p> </p> | + | <p> </p> |
- | </div> | + | </div> |
- | </div> | + | </div> |
<!-- 1010101010101010101010101010 --> | <!-- 1010101010101010101010101010 --> | ||
- | + | <div class="inner" id="week11"> | |
- | <script type="text/javascript"> | + | <div class="block" id="summ"> |
+ | <h3 style="color:white;">Data&protocol</h3> | ||
+ | <hr/> | ||
+ | <div class="block" id="monday"> | ||
+ | <h3>Monday</h3> | ||
+ | <hr/> | ||
+ | <p>WEEK11</p> | ||
+ | </div> | ||
+ | <div class="block" id="tuesday"> | ||
+ | <h3>Tuesday</h3> | ||
+ | <hr/> | ||
+ | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> | ||
+ | <p> </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | <!-- 11 11 11 11 11 11 11 11 11 --> | ||
+ | <div class="inner" id="week12"> | ||
+ | <div class="block" id="summ"> | ||
+ | <h3 style="color:white;">Data&protocol</h3> | ||
+ | <hr/> | ||
+ | <div class="block" id="monday"> | ||
+ | <h3>Monday</h3> | ||
+ | <hr/> | ||
+ | <p>WEEK12</p> | ||
+ | </div> | ||
+ | <div class="block" id="tuesday"> | ||
+ | <h3>Tuesday</h3> | ||
+ | <hr/> | ||
+ | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> | ||
+ | <p> </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | <!-- 12 12 12 12 12 12 12 12 12 12 12 --> | ||
+ | <div class="inner" id="afweek12"> | ||
+ | <div class="block" id="summ"> | ||
+ | <h3 style="color:white;">Data&protocol</h3> | ||
+ | <hr/> | ||
+ | <div class="block" id="monday"> | ||
+ | <h3>Monday</h3> | ||
+ | <hr/> | ||
+ | <p>WEEK13</p> | ||
+ | </div> | ||
+ | <div class="block" id="tuesday"> | ||
+ | <h3>Tuesday</h3> | ||
+ | <hr/> | ||
+ | <p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div> | ||
+ | <p> </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | <!-- 13 13 13 13 13 13 13 13 13 13 13 13 --> | ||
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- | + | ||
- | + | ||
- | </div></div><p style ="clear:both"></p> | + | </div></div><p style ="clear:both"></p> |
- | </div> | + | </div> |
- | <div class="footer" id="footer"><a href="https://2011.igem.org/"> iGem 2011 Home Page</a> | + | <div class="footer" id="footer"><a href="https://2011.igem.org/"> iGem 2011 Home Page</a> |
- | <a href="http://ung.igem.org/Special:Upload/"> Upload Files</a> | + | <a href="http://ung.igem.org/Special:Upload/"> Upload Files</a> |
- | <a href="https://2011.igem.org/wiki/index.php?title=Template:Zjucss_main&action=edit">Edit CSS</a> | + | <a href="https://2011.igem.org/wiki/index.php?title=Template:Zjucss_main&action=edit">Edit CSS</a> |
- | <a href="http://ung.igem.org/Team_Wikis?year=2011"> Team Wikis</a> | + | <a href="http://ung.igem.org/Team_Wikis?year=2011"> Team Wikis</a> |
- | <a href="mailto:zjuigem@gmail.com?"><strong>Contact Us</strong></a> | + | <a href="mailto:zjuigem@gmail.com?"><strong>Contact Us</strong></a> |
- | </div> | + | </div> |
- | </div> | + | </div> |
- | </body> | + | </body> |
</html> | </html> |
Revision as of 15:51, 26 September 2011
<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
Week1
Day | Note | ||
Jul.4th Monday |
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Jul 5th Tuesday | |||
Jul.6th Wednesday |
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Jul.7th Thursday |
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Jul.8th Friday |
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Jul.9th Saturday |
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Jul.10th Sunday |
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Data&protocol
Monday
There is no data in first week
Week2
Day | Note | |||||
Jul.11th Monday |
|
|||||
Jul 12th Tuesday |
|
|||||
Jul.13th Wednesday |
|
|||||
Jul.14th Thursday |
|
|||||
Jul.15th Friday |
|
|||||
Jul.16th Saturday |
|
|||||
Jul.17th Sunday |
|
Data&protocol
Jul.13th Wednesday
Systems of restriction digestion with EcoRI and PstI
Plasmid | 1μL (>100ng) |
EcoRI | 1μL |
PstI | 1μL |
10×Buffer Tango | 2μL |
ddH2O | 15μL |
Temperature grad
56℃,57.4℃,60.2℃,62.9℃,64.3℃,65.8℃
PCR system (test the Tm of the primers CP1&CS, NP&NS)
10×Buffer | 2μL |
dNTPs | 0.5μL |
primers CP1 (NP) | 1μL |
primers CS (NS) | 1μL |
Template | 1μL |
rTaq | 0.2μL |
H2O | 14.5μL |
Total | 20μL |
Jul.14th Thursday
PCR systems(test the Tm of the primers for YFP,RFP,tetR,Vgb and nirB )
10×Buffer | 2μL |
dNTPs | 0.5μL |
primers F | 0.5μL |
primers R | 0.5μL |
Template | 1μL |
Taq | 0.2μL |
H2O | 15.5μL |
Total | 20μL |
Temperature grad and the results:
vgb&nirB pcr test:
Week3
Day | Note | ||||
Jul.18th Monday |
|
||||
Jul 19th Tuesday |
|
||||
Jul.20th Wednesday |
|
||||
Jul.21th Thursday |
|
||||
Jul.22th Friday |
|
||||
Jul.23th Saturday |
|
||||
Jul.24th Sunday |
|
Data&protocol
Monday
WEEK3
Tuesday
bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb
Week4
Day | Note | ||||
Jul.25th Monday |
|
||||
Jul. 26th Tuesday |
|
||||
Jul.27th Wednesday |
|
||||
Jul.28th Thursday |
|
||||
Jul.29th Friday |
|
||||
Jul.30th Saturday | |||||
Jul.31th Sunday |
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Data&protocol
Monday
WEEK3
Tuesday
bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb
Week5
Day | Note | ||||
Aug.1st Monday |
|
||||
Aug.2nd Tuesday |
|
||||
Aug.3rd Wednesday |
|
||||
Aug.4th Thursday |
|
||||
Aug.5th Friday |
|
||||
Aug.6th Saturday | |||||
Aug.7th Sunday |
|
Data&protocol
Monday
WEEK3
Tuesday
bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb
Week6
Day | Note | ||||
Aut.8th Monday |
|
||||
Aug.9th Tuesday |
|
||||
Aug.10th Wednesday |
|
||||
Aug.11th Thursday |
|
||||
Aug.12th Friday |
|
||||
Aug.13th Saturday |
|
||||
Aug.14th Sunday |
|
Data&protocol
Monday
WEEK3
Tuesday
bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb
Data&protocol
Monday
WEEK7
Tuesday
bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb
Data&protocol
Monday
WEEK8
Tuesday
bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb
Data&protocol
Monday
WEEK9
Tuesday
bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb
Data&protocol
Monday
WEEK10
Tuesday
bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb
Data&protocol
Monday
WEEK11
Tuesday
bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb
Data&protocol
Monday
WEEK12
Tuesday
bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb
Data&protocol
Monday
WEEK13
Tuesday
bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb