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

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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Introduction">Overview</a></li>
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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/H_Stem">H-Stem System</a></li>
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                                                        <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Project/H_Stem#Modeling">Modelling</a></li>
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                                                            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034/Method">Method</a></li>
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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length/Overview">Length Variation</a></li>
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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Materials">Extra Materials</a></li>
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<span class="menu-list">Policy and Practice</span>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Discussion#Background">Background</a></li>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Discussion#Estimation">Estimation</a></li>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/About_Us#Acknowledgements">Acknowledgements</a></li>
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{{Team:HokkaidoU_Japan/JS}}

Revision as of 11:15, 14 October 2014

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 & 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!

Retrieved from "http://2014.igem.org/Team:HokkaidoU_Japan/Notebook/Pre_experiment/mRFP-expression-construct"