Team:HokkaidoU Japan/Notebook/Promoter RandomPromtoer-B0034-mRFP1-B0015

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<h2>RandomPromtoer-B0034-mRFP1-B0015</h2>
<div class="workflow-container">
  <ul class="steps">
    <li class="step start">
      <div class="step-tile"><h2>Start</h2></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>Tue Jun 11</time> </li> </ul> <div class="step-recipe"> B0034-mRFP1-B0015<br> 100up, 200dn, KOD Plus Neo </div> </div> </div> </li>
<li class="step"> <div class="step-tile"> <h2 class="step-title">Get K1084007 from stock</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Tue Jun 11</time> </li> </ul> <div class="step-recipe"> </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>Wed Jun 12</time> </li> </ul> <div class="step-recipe"> B0034-mRFP1-B0015 </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>Wed Jun 12</time> </li> </ul> <div class="step-recipe"> Cut K1084007 with SpeI, PstI (using 10&times;H buffer)<br> Cut B0034-mRFP1-B0015 with XbaI, PstI (using 10&times;M buffer) </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>Thu Jun 13</time> </li> </ul> <div class="step-recipe"> Ligate K1084007 with B0034-mRFP1-B0015 </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>Thu Jun 13</time> </li> </ul> <div class="step-recipe"> K1084007-B0034-mRFP1-B0015<br> 1&micro;L DNA to DH5&alpha; </div> </div> </div> </li>
<li class="step"> <div class="step-tile"> <h2 class="step-title">Colony PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Mon Jun 17</time> </li> </ul> <div class="step-recipe"> K1084007-B0034-mRFP1-B0015 </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>Tue Jun 18 and Wed Jun 19</time> </li> </ul> <div class="step-recipe"> K1084007(colony)<br> ranori-f, 200dn, KOD FX Neo<br> We got random promter.<br><br> B0034-LacZ&alpha;-B0015(colony)<br> 100up, 200dn, KOD FX 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>Thu Jun 20</time> </li> </ul> <div class="step-recipe"> Cut random promoter with PstI, SpeI (using 10&times;H buffer)<br> Cut B0034-LacZ&alpha;-B0015 with XbaI, PstI (using 10&times;M buffer) </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>Thu Jun 20</time> </li> </ul> <div class="step-recipe"> Ligate random promoter with B0034-LacZ&alpha;-B0015 </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>Thu Jun 20</time> </li> </ul> <div class="step-recipe"> 1&micro;L DNA to JM109<br> to X-Gal plate </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>Fri Jun 21</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-LacZ&alpha;-B0015<br> to X-Gal 50mg/mL, X-Gal 5mg/mL and LBC<br> We got some blue colonies with different levels of blue. </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 22</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-LacZ&alpha;-B0015 &times;10 samples<br> random promoter-B0034-mRFP1-B0015 &times;9 samples<br> K1084007-B0034-mRFP1-B0015 </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 23</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-LacZ&alpha;-B0015 &times;10 samples<br> random promoter-B0034-mRFP1-B0015 &times;9 samples<br> K1084007-B0034-mRFP1-B0015 </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>Wed Jun 26</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-mRFP1-B0015<br> K1084007-B0034-mRFP1-B0015<br> random promoter-B0034-mRFP1-B0015 &times;20 samples </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>Thu Jun 27</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-mRFP1-B0015 &times;2 samples<br> random promoter-B0034-LacZ&alpha;-B0015 &times;2 samples<br> 100up, 200dn, KOD FX Neo </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 Jun 28</time> </li> </ul> <div class="step-recipe"> J23112, J23103, J23117, J23106, J23111, J23100<br> 100up, 200dn, KOD FX 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>Fri Jun 28</time> </li> </ul> <div class="step-recipe"> Cut (J23112, J23103, J23117, J23106, J23111, J23100) with EcoRI, PstI (using 10&times;H buffer)<br> Cut B0034-mRFP1-B0015 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>Fri Jun 28</time> </li> </ul> <div class="step-recipe"> B0034-mRFP1-B0015 </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>Sat Jun 29</time> </li> </ul> <div class="step-recipe"> Ligate (J23112, J23103, J23117, J23106, J23111, J23100) with B0034-mRFP1-B0015 </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>Sat Jun 29 and Fri Jul 5</time> </li> </ul> <div class="step-recipe"> (J23112, J23103, J23117, J23106, J23111, J23100)-B0034-mRFP1-B0015<br> 1&micro;L DNA to JM109<br> pre culture </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>Mon Jul 1</time> </li> </ul> <div class="step-recipe"> (J23112, J23103, J23117, J23106, J23111, J23100) with B0034-mRFP1-B0015<br> We picked up some colonies to a X-Gal LBC plate. We confirmed that colonies were white. </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>Sat Jul 6</time> </li> </ul> <div class="step-recipe"> R0040-B0034-mRFP1-B0015 </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>Mon Jul 8</time> </li> </ul> <div class="step-recipe"> (J23112, J23103, J23117, J23106, J23111, J23100, R0040, R0010)-B0034-mRFP1-B0015 </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>Thu Jul 11</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-mRFP1-B0015<br> 1&micro;L DNA to JM109 to X-Gal plate </div> </div> </div> </li>
<li class="step"> <div class="step-tile"> <h2 class="step-title">PCR & PCR purification</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Mon Jul 15</time> </li> </ul> <div class="step-recipe"> J23112, J23103, J23117(colony)<br> 100up, 200dn, KOD FX 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 Jul 15</time> </li> </ul> <div class="step-recipe"> Cut J23112, J23103, J23117 with EcoRI, PstI (using 10&times;H buffer)<br> Cut random promoter-B0034-LacZ&alpha;-B0015 on pSB1C3 with EcoRI, PstI (using 10&times;M buffer) </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>Tue Jul 16</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>Tue Jul 16</time> </li> </ul> <div class="step-recipe"> Ligate (J23112, J23103, J23117) with pSB1C3 </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 Jul 16 and Fri Jul 19</time> </li> </ul> <div class="step-recipe"> 1&micro;L DNA to JM109 to 10 X-Gal plates </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>Fri Jul 26</time> </li> </ul> <div class="step-recipe"> We cultured some colonies from Jul 19 and 23 plates to X-Gal plate. </div> </div> </div> </li>
<li class="step"> <div class="step-tile"> <h2 class="step-title">Assay</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Wed Aug 7</time> </li> </ul> <div class="step-recipe"> Assay for RFP expression </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>Thu Aug 8, Fri Aug 9 and Sat Aug 10</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-mRFP1-B0015 </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>Sat Aug 10</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-mRFP1-B0015(cultured Aug 8) </div> </div> </div> </li>
    <li class="step complete">
      <div class="step-tile"><h2>Complete!</h2></div>
    </li>
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