Team:HokkaidoU Japan/Notebook/RBS SD2,4,6

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<h2>SD2, 4, 6</h2>
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       <div class="step-tile"><h2>Start</h2></div>
       <div class="step-tile"><h2>Start</h2></div>
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<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"> LacZ&alpha;-B0015 100up 200dn </div> </div> </div> </li>
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<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-LacZ&alpha;-B0015 / SD8 100up SD(2/4/6) mutation revers </div> </div> </div> </li>
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<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>
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<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 LacZ&alpha;-B-0015 EcoRI and XbaI (using 10&times;M buffer) , PCR products EcoRI and SpeI (using 10&times;H buffer) </div> </div> </div> </li>
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<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>
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<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"> LacZ&alpha;-B-0015 </div> </div> </div> </li>
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<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"> LacZ&alpha;-B-0015 </div> </div> </div> </li>
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<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 LacZ&alpha;-B0015<br> Used X-cut-vector for inserts with R0040, and S-cut-vector for insrts without R0040 </div> </div> </div> </li>
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<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)LacZ&alpha;-B0015 / SD(6/4/2)<br> each of 4&micro;L DNA to DH5&alpha; </div> </div> </div> </li>
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<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!