Team:HokkaidoU Japan/Notebook/Promoter WildPromoter-I732020

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<h2>WildPromoter-I732020</h2>
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       <div class="step-tile"><h2>Start</h2></div>
       <div class="step-tile"><h2>Start</h2></div>
     </li>
     </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>Fri Jun 7</time> </li> </ul> <div class="step-recipe"> J23112, J23103, J23117, J23106, J23111 from distribution kit<br> each of 1&micro;L DNA to DH5&alpha; </div> </div> </div> </li>
 +
<li class="step"> <div class="step-tile"> <h2 class="step-title">Culture</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sat Jun 8 and Fri Jun 21</time> </li> </ul> <div class="step-recipe"> J23112, J23103, J23117, J23106, J23111<br> I732020 from our stock </div> </div> </div> </li>
 +
<li class="step"> <div class="step-tile"> <h2 class="step-title">Mini-prep</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun Jun 9 and Sat Jun 22</time> </li> </ul> <div class="step-recipe"> J23112, J23103, J23117, J23106, J23111, I732020 </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>Wed Jun 26</time> </li> </ul> <div class="step-recipe"> I732020<br> 100up, 200dn, KOD Plus Neo </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>Mon Jun 10, Sat Jun 22 and Wed Jun 26</time> </li> </ul> <div class="step-recipe"> Cut (J23112, J23103, J23117, J23106, J23111) with SpeI, PstI (using 10&times;H buffer)<br> Cut I732020 with XbaI, PstI (using 10&times;M buffer) </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>Tue Jun 11 and Mon Jun 24</time> </li> </ul> <div class="step-recipe"> J23112, J23103, J23117, J23106, J23111 </div> </div> </div> </li>
 +
<li class="step"> <div class="step-tile"> <h2 class="step-title">Antarctic phosphatase</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun Jun 30</time> </li> </ul> <div class="step-recipe"> J23112, J23103, J23117, J23106, J23111 </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>Tue Jun 11, Mon Jun 24 and Sun Jun 30</time> </li> </ul> <div class="step-recipe"> Ligate (J23112, J23103, J23117, J23106, J23111) with I732020 </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>Tue Jun 11, Mon Jun 24 and Sun Jun 30</time> </li> </ul> <div class="step-recipe"> (J23112, J23103, J23117, J23106, J23111)-I732020<br> each of 1&micro;L DNA to DH5&alpha; </div> </div> </div> </li>
 +
<li class="step"> <div class="step-tile"> <h2 class="step-title">Colony PCR & Culture</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun Jun 30</time> </li> </ul> <div class="step-recipe"> (J23112, J23103, J23117, J23106, J23111)-I732020 </div> </div> </div> </li>
 +
<li class="step"> <div class="step-tile"> <h2 class="step-title">Mini-prep</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Mon Jul 1</time> </li> </ul> <div class="step-recipe"> (J23112, J23103, J23117, J23106, J23111)-I732020 </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>Fri Jul 5</time> </li> </ul> <div class="step-recipe"> (J23112, J23103, J23117, J23106, J23111)-I732020<br> EX-f, PS-r, KOD FX NEO </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 Jul 6</time> </li> </ul> <div class="step-recipe"> (J23112, J23103, J23117, J23106, J23111)-I732020 </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 Jul 6</time> </li> </ul> <div class="step-recipe"> Cut K1084007 on pSB1C3 with EcoRI, PstI (using 10&times;H buffer)<br> Cut (J23112, J23103, J23117, J23106, J23111)-I732020 with EcoRI, PstI (using 10&times;H buffer) </div> </div> </div> </li>
 +
<li class="step"> <div class="step-tile"> <h2 class="step-title">Antarctic phophatase</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun Jul 7</time> </li> </ul> <div class="step-recipe"> pSB1C3 </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 Jul 7</time> </li> </ul> <div class="step-recipe"> Ligate pSB1C3 with (J23112, J23103, J23117, J23106, J23111)-I732020 </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 Jul 7</time> </li> </ul> <div class="step-recipe"> (J23112, J23103, J23117, J23106, J23111)-I732020 </div> </div> </div> </li>
     <li class="step complete">
     <li class="step complete">
       <div class="step-tile"><h2>Complete!</h2></div>
       <div class="step-tile"><h2>Complete!</h2></div>

Revision as of 06:59, 27 September 2013

Maestro E.coli

Notebook

WildPromoter-I732020

  • Start

  • Transformation

    J23112, J23103, J23117, J23106, J23111 from distribution kit
    each of 1µL DNA to DH5α
  • Culture

    J23112, J23103, J23117, J23106, J23111
    I732020 from our stock
  • Mini-prep

    J23112, J23103, J23117, J23106, J23111, I732020
  • PCR

    I732020
    100up, 200dn, KOD Plus Neo
  • Digestion

    Cut (J23112, J23103, J23117, J23106, J23111) with SpeI, PstI (using 10×H buffer)
    Cut I732020 with XbaI, PstI (using 10×M buffer)
  • Gel extraction

    J23112, J23103, J23117, J23106, J23111
  • Antarctic phosphatase

    J23112, J23103, J23117, J23106, J23111
  • Ligation

    Ligate (J23112, J23103, J23117, J23106, J23111) with I732020
  • Transformation

    (J23112, J23103, J23117, J23106, J23111)-I732020
    each of 1µL DNA to DH5α
  • Colony PCR & Culture

    (J23112, J23103, J23117, J23106, J23111)-I732020
  • Mini-prep

    (J23112, J23103, J23117, J23106, J23111)-I732020
  • PCR

    (J23112, J23103, J23117, J23106, J23111)-I732020
    EX-f, PS-r, KOD FX NEO
  • PCR purification

    (J23112, J23103, J23117, J23106, J23111)-I732020
  • Digestion

    Cut K1084007 on pSB1C3 with EcoRI, PstI (using 10×H buffer)
    Cut (J23112, J23103, J23117, J23106, J23111)-I732020 with EcoRI, PstI (using 10×H buffer)
  • Antarctic phophatase

    pSB1C3
  • Ligation

    Ligate pSB1C3 with (J23112, J23103, J23117, J23106, J23111)-I732020
  • Transformation

    (J23112, J23103, J23117, J23106, J23111)-I732020
  • Complete!