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

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<div class="wrapper">
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   <div id="hokkaidou-contents">
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<h2> mRFP Expression Vector Construction</h2>
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<h2> mRFP Expression Plasmid Construction</h2>
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      <div class="step-recipe"> pSB6A1
      <div class="step-recipe"> pSB6A1
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1&micro;L DNA to JM109</div>
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1&micro;L to JM109</div>
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      <div class="step-recipe"> R0040-B0034-E1010-B0015 from our stock
      <div class="step-recipe"> R0040-B0034-E1010-B0015 from our stock
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100up 200dn</div>
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100up_F Tm:70.4&#8451; & 200dn_R Tm:66.6&#8451;
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        KOD plus Neo</div>
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      </ul>
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      <div class="step-recipe"> Cut R0040-B0034-E1010-B0015 with EcoRI, PstI (using 10&times;H buffer)
      <div class="step-recipe"> Cut R0040-B0034-E1010-B0015 with EcoRI, PstI (using 10&times;H buffer)
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Cut pSB6A1 with EcoRI, PstI (using 10&times;H buffer)</div>
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Cut pSB6A1 with EcoRI, PstI (using 10&times;H buffer)
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      <div class="step-recipe"> R0040-B0034-E1010-B0015
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pSB6A1</div>
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pSB6A1
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  <time>Sat Aug 16</time>
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pSB6A1
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      <div class="step-recipe"> R0040-B0034-E1010-B0015</div>
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      <div class="step-recipe"> Ligate R0040-B0034-E1010-B0015 with pSB6A1</div>
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      <div class="step-recipe"> Ligate R0040-B0034-E1010-B0015 with pSB6A1
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      <div class="step-recipe"> R0040-B0034-E1010-B0015 on pSB6A1
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      <div class="step-recipe"> R0040-B0034-E1010-B0015(on pSB6A1)
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5&micro;L DNA to DH5&alpha; Turbo</div>
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5&micro;L to DH5&alpha; Turbo
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      <div class="step-recipe"> R0040-B0034-E1010-B0015 on pSB6A1</div>
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      <div class="step-recipe"> R0040-B0034-E1010-B0015(on pSB6A1)
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      <div class="step-recipe"> R0040-B0034-E1010-B0015 on pSB6A1</div>
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      <div class="step-recipe">R0040-B0034-E1010-B0015(on pSB6A1)</div>
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{{Team:HokkaidoU_Japan/JS}}

Latest revision as of 15:24, 9 September 2015

Notebook
Lab Documents

mRFP Expression Plasmid Construction

  • Start

  • Get genes

    pSB6A1

  • Transformation

    pSB6A1 1µL to JM109

  • Liquid Culture

    pSB6A1

  • Mini-prep

    pSB6A1

  • PCR & PCR purification

    R0040-B0034-E1010-B0015 from our stock 100up_F Tm:70.4℃ & 200dn_R Tm:66.6℃
    KOD plus Neo

  • 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 to DH5α Turbo

  • Liquid Culture

    R0040-B0034-E1010-B0015(on pSB6A1)

  • Mini-prep

    R0040-B0034-E1010-B0015(on pSB6A1)

  • Complete!

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