Team:HokkaidoU Japan/Notebook/RBS SD2,4,6
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- | <h2></h2> | + | <h2>SD2, 4, 6</h2> |
<div class="workflow-container"> | <div class="workflow-container"> | ||
<ul class="steps"> | <ul class="steps"> | ||
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<div class="step-tile"><h2>Start</h2></div> | <div class="step-tile"><h2>Start</h2></div> | ||
</li> | </li> | ||
- | + | <li class="step"> <div class="step-tile"> <h2 class="step-title">PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sat May 4</time> </li> </ul> <div class="step-recipe"> LacZα-B0015 100up 200dn </div> </div> </div> </li> | |
+ | <li class="step"> <div class="step-tile"> <h2 class="step-title">PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sat May 11</time> </li> </ul> <div class="step-recipe"> R0040-SD8-LacZα-B0015 / SD8 100up SD(2/4/6) mutation revers </div> </div> </div> </li> | ||
+ | <li class="step"> <div class="step-tile"> <h2 class="step-title">PCR purification</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sat May 11</time> </li> </ul> <div class="step-recipe"> R0040-SD(6/4/2) / SD(6/4/2) </div> </div> </div> </li> | ||
+ | <li class="step"> <div class="step-tile"> <h2 class="step-title">Digestion</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sat May 11</time> </li> </ul> <div class="step-recipe"> Cut LacZα-B-0015 EcoRI and XbaI (using 10×M buffer) , PCR products EcoRI and SpeI (using 10×H buffer) </div> </div> </div> </li> | ||
+ | <li class="step"> <div class="step-tile"> <h2 class="step-title">Ethanol precipitation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun May 12</time> </li> </ul> <div class="step-recipe"> R0040-SD(6/4/2) / SD(6/4/2)<br> </div> </div> </div> </li> | ||
+ | <li class="step"> <div class="step-tile"> <h2 class="step-title">Gel extraction</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun May 12</time> </li> </ul> <div class="step-recipe"> LacZα-B-0015 </div> </div> </div> </li> | ||
+ | <li class="step"> <div class="step-tile"> <h2 class="step-title">Ethanol precipitation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun May 12</time> </li> </ul> <div class="step-recipe"> LacZα-B-0015 </div> </div> </div> </li> | ||
+ | <li class="step"> <div class="step-tile"> <h2 class="step-title">Ligation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun May 12</time> </li> </ul> <div class="step-recipe"> Ligate R0040-SD(6/4/2) / SD(6/4/2) with LacZα-B0015<br> Used X-cut-vector for inserts with R0040, and S-cut-vector for insrts without R0040 </div> </div> </div> </li> | ||
+ | <li class="step"> <div class="step-tile"> <h2 class="step-title">Transformation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun May 12</time> </li> </ul> <div class="step-recipe"> R0040-SD(6/4/2)LacZα-B0015 / SD(6/4/2)<br> each of 4µL DNA to DH5α </div> </div> </div> </li> | ||
+ | <li class="step"> <div class="step-tile"> <h2 class="step-title">Sequence</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Fri Sep 6</time> </li> </ul> <div class="step-recipe"> SD(6/4/2) on pSB1C3 </div> </div> </div> </li> | ||
<li class="step complete"> | <li class="step complete"> |
Latest revision as of 16:37, 27 September 2013
Maestro E.coli
Notebook
SD2, 4, 6
-
Start
-
PCR
LacZα-B0015 100up 200dn -
PCR
R0040-SD8-LacZα-B0015 / SD8 100up SD(2/4/6) mutation revers -
PCR purification
R0040-SD(6/4/2) / SD(6/4/2) -
Digestion
Cut LacZα-B-0015 EcoRI and XbaI (using 10×M buffer) , PCR products EcoRI and SpeI (using 10×H buffer) -
Ethanol precipitation
R0040-SD(6/4/2) / SD(6/4/2)
-
Gel extraction
LacZα-B-0015 -
Ethanol precipitation
LacZα-B-0015 -
Ligation
Ligate R0040-SD(6/4/2) / SD(6/4/2) with LacZα-B0015
Used X-cut-vector for inserts with R0040, and S-cut-vector for insrts without R0040 -
Transformation
R0040-SD(6/4/2)LacZα-B0015 / SD(6/4/2)
each of 4µL DNA to DH5α -
Sequence
SD(6/4/2) on pSB1C3 -
Complete!