Team:HokkaidoU Japan/Notebook/Promoter K1084001~010-B0034-lacZ-B0015

From 2013.igem.org

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      <h1 id="common-header-title">Maestro E.coli</h1>
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      <h2 id="common-header-subtitle">Notebook</h2>
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<h2>K1084001-010-B0034-lacZ-B0015</h2>
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<div class="workflow-container">
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  <ul class="steps">
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    <li class="step start">
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      <div class="step-tile"><h2>Start</h2></div>
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    </li>
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<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>Sun Aug 11, Mon Sep 2</time> </li> </ul> <div class="step-recipe"> K1084007-B0034-mRFP1-B0015<br> 100b up, random-oripro-r, KOD plus Neo </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>Mon Aug 12, Tue Sep 3</time> </li> </ul> <div class="step-recipe"> Cut random promoter(PCR product) with EcoRI, SpeI(using 10xH buffer)<br> Cut K1084007-B0034-mRFP1-B0015 on pSB1C3 with EcoRI, SpeI(using 10xH buffer) </div> </div> </div> </li>
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<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>Mon Aug 12, Wed Aug 14, Tue Sep 3</time> </li> </ul> <div class="step-recipe"> pSB1C3 </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>Mon Aug 12, Wed Aug 14, Tue Sep 3</time> </li> </ul> <div class="step-recipe"> Ligate random promoter with pSB1C3 </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>Mon Aug 12, Wed Aug 14, Tue Sep 3</time> </li> </ul> <div class="step-recipe"> random promoter on pSB1C3<br> 1&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">Colony PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu Aug 15, Thu Sep 5</time> </li> </ul> <div class="step-recipe"> random promoter on pSB1C3 </div> </div> </div> </li>
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<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 15</time> </li> </ul> <div class="step-recipe"> K1084007-B0034-mRFP1-B0015 on pSB1C3 </div> </div> </div> </li>
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<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>Fri Aug 16</time> </li> </ul> <div class="step-recipe"> K1084007-B0034-mRFP1-B0015 on pSB1C3 </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>Fri Aug 16</time> </li> </ul> <div class="step-recipe"> K1084007:100b up, oripro-r, KOD plus Neo<br> pSB1C3:Suf-I-F, XE-R, KOD plus Neo </div> </div> </div> </li>
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<li class="step"> <div class="step-tile"> <h2 class="step-title">EtOH precipitation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sat Aug 17</time> </li> </ul> <div class="step-recipe"> K1084007, pSB1C3 </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 Aug 17</time> </li> </ul> <div class="step-recipe"> Cut K1084007 with EcoRI, SpeI(using 10xH buffer) <br> Cut pSB1C3 with EcoRI, SpeI(using 10xH buffer) </div> </div> </div> </li>
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<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 Aug 18</time> </li> </ul> <div class="step-recipe"> K1084007 </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 Aug 18</time> </li> </ul> <div class="step-recipe"> Ligate K1084007 with pSB1C3 </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 Aug 18</time> </li> </ul> <div class="step-recipe"> K1084007 on pSB1C3<br> 1&micro;L DNA to JM109 </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>Mon Aug 19</time> </li> </ul> <div class="step-recipe"> random promoter on pSB1C3<br> 1&micro;L DNA to JM109 </div> </div> </div> </li>
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<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>Wed Aug 21, Fri Sep 6</time> </li> </ul> <div class="step-recipe"> random promoter on pSB1C3 </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>Thu Aug 22</time> </li> </ul> <div class="step-recipe"> Ligate random promoter with pSB1C3 </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>Thu Aug 22</time> </li> </ul> <div class="step-recipe"> random promoter on pSB1C3 </div> </div> </div> </li>
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<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>Wed Aug 28</time> </li> </ul> <div class="step-recipe"> random promoter on pSB1C3 </div> </div> </div> </li>
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<li class="step"> <div class="step-tile"> <h2 class="step-title">Sequencing</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Fri Aug 30, Sun Sep 1, Mon Sep 2, Fri Sep 6, Mon Sep 9</time> </li> </ul> <div class="step-recipe"> random promoter on pSB1C3 </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>Fri Sep 6</time> </li> </ul> <div class="step-recipe"> Cut I732020 with XbaI, PstI(using 10xM buffer)<br> Cut R0040-B0034-mRFP1-B0015 on pSB1C3 with EcoRI, PstI(using 10xH buffer) </div> </div> </div> </li>
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<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 Sep 15</time> </li> </ul> <div class="step-recipe"> pSB1C3 </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 Sep 15</time> </li> </ul> <div class="step-recipe"> Ligate random promoter with B0034-lacZ&alpha;-B0015, pSB1C3 </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 Sep 15</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZ&alpha;-B0015 on pSB1C3<br> 5&micro;L DNA to JM109 </div> </div> </div> </li>
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<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>Tue Sep 17</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZ&alpha;-B0015 on pSB1C3 </div> </div> </div> </li>
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<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 Sep 18, Thu Sep 19</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZ&alpha;-B0015 on pSB1C3<br> <br> random promoter:K1084001~010 </div> </div> </div> </li>
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<li class="step"> <div class="step-tile"> <h2 class="step-title">Sequencing</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu Sep 19</time> </li> </ul> <div class="step-recipe"> K1084001~010-B0034-lacZ&alpha;-B0015<br> 200b dn </div> </div> </div> </li>
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<li class="step"> <div class="step-tile"> <h2 class="step-title">&beta;-galactosidase assay</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Fri Sep 20, Thu Sep 26</time> </li> </ul> <div class="step-recipe"> K1084001~010-B0034-lacZ&alpha;-B0015 on pSB1C3 </div> </div> </div> </li>
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    <li class="step complete">
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      <div class="step-tile"><h2>Complete!</h2></div>
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    </li>
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  </ul>
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Latest revision as of 07:03, 27 September 2013

Maestro E.coli

Notebook

K1084001-010-B0034-lacZ-B0015

  • Start

  • PCR & PCR purification

    K1084007-B0034-mRFP1-B0015
    100b up, random-oripro-r, KOD plus Neo
  • Digestion

    Cut random promoter(PCR product) with EcoRI, SpeI(using 10xH buffer)
    Cut K1084007-B0034-mRFP1-B0015 on pSB1C3 with EcoRI, SpeI(using 10xH buffer)
  • Antarctic phosphatase

    pSB1C3
  • Ligation

    Ligate random promoter with pSB1C3
  • Transformation

    random promoter on pSB1C3
    1µL DNA to DH5α
  • Colony PCR

    random promoter on pSB1C3
  • Culture

    K1084007-B0034-mRFP1-B0015 on pSB1C3
  • Mini-prep

    K1084007-B0034-mRFP1-B0015 on pSB1C3
  • PCR

    K1084007:100b up, oripro-r, KOD plus Neo
    pSB1C3:Suf-I-F, XE-R, KOD plus Neo
  • EtOH precipitation

    K1084007, pSB1C3
  • Digestion

    Cut K1084007 with EcoRI, SpeI(using 10xH buffer)
    Cut pSB1C3 with EcoRI, SpeI(using 10xH buffer)
  • Antarctic phosphatase

    K1084007
  • Ligation

    Ligate K1084007 with pSB1C3
  • Transformation

    K1084007 on pSB1C3
    1µL DNA to JM109
  • Transformation

    random promoter on pSB1C3
    1µL DNA to JM109
  • Colony PCR

    random promoter on pSB1C3
  • Ligation

    Ligate random promoter with pSB1C3
  • Transformation

    random promoter on pSB1C3
  • Colony PCR

    random promoter on pSB1C3
  • Sequencing

    random promoter on pSB1C3
  • Digestion

    Cut I732020 with XbaI, PstI(using 10xM buffer)
    Cut R0040-B0034-mRFP1-B0015 on pSB1C3 with EcoRI, PstI(using 10xH buffer)
  • Antarctic phosphatase

    pSB1C3
  • Ligation

    Ligate random promoter with B0034-lacZα-B0015, pSB1C3
  • Transformation

    random promoter-B0034-lacZα-B0015 on pSB1C3
    5µL DNA to JM109
  • Colony PCR

    random promoter-B0034-lacZα-B0015 on pSB1C3
  • Culture

    random promoter-B0034-lacZα-B0015 on pSB1C3

    random promoter:K1084001~010
  • Sequencing

    K1084001~010-B0034-lacZα-B0015
    200b dn
  • β-galactosidase assay

    K1084001~010-B0034-lacZα-B0015 on pSB1C3
  • Complete!