Team:HokkaidoU Japan/Notebook/Pre experiment/mRFP-expression-construct

From 2014.igem.org

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  <h2 class="step-title">Gel Extraction & Ethanol precipetation</h2>
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  <h2 class="step-title">Gel Extraction</h2>
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      <ul class="step-detail-trials">
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        <li>
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  <time>Sat Aug 16</time>
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      </ul>
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      <div class="step-recipe"> R0040-B0034-E1010-B0015
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pSB6A1</div>
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  <h2 class="step-title">Ethanol precipetation</h2>
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Revision as of 13:09, 17 October 2014

hogehoge

mRFP Expression Plasmid Construction

  • Start

  • Get genes

    pSB6A1
  • Transformation

    pSB6A1 1µL DNA to JM109
  • Liquid Culture

    pSB6A1
  • Mini-prep

    pSB6A1
  • PCR & PCR purification

    R0040-B0034-E1010-B0015 from our stock 100up 200dn
  • Digestion

    Cut R0040-B0034-E1010-B0015 with EcoRI, PstI (using 10×H buffer) Cut pSB6A1 with EcoRI, PstI (using 10×H buffer)
  • Gel Extraction

    R0040-B0034-E1010-B0015 pSB6A1
  • Ethanol precipetation

    R0040-B0034-E1010-B0015 pSB6A1
  • Dephosphorylation

    R0040-B0034-E1010-B0015
  • Ligation

    Ligate R0040-B0034-E1010-B0015 with pSB6A1
  • Transformation

    R0040-B0034-E1010-B0015 on pSB6A1 5µL DNA to DH5α Turbo
  • Liquid Culture

    R0040-B0034-E1010-B0015 on pSB6A1
  • Mini-prep

    R0040-B0034-E1010-B0015 on pSB6A1
  • Complete!