Team:HokkaidoU Japan/Notebook/Length/as90 120

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Anti-sense 90, 120 construction
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Anti-sense 90, 120 Construction
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Revision as of 11:39, 12 October 2014

   
    

Anti-sense 90, 120 Construction

  • Start

  • PCR & PCR purification

    R0040-B0034-E1010-B0015 as RBS XhoI, 90bp as mRFP Nco as RBS XhoI, 120bp as mRFP Nco
  • Digestion

    Cut as 90, as120, H-stem with NcoI, XhoI(using 10×Cut Smart)
  • Ethanol precipitation

    as90, as120
  • Gel Extraction

    H-stem
  • Dephophorylation

    H-stem
  • Ligation

    Ligate as90, as120 with H-stem
  • Transformation

    as90, as120 on H-stem 5µL DNA to DH5α Turbo
  • Colony PCR

    as90, as120 on H-stem
  • Liquid Culture

    as90, as120 on H-stem
  • Mini-prep

    as90, as120 on H-stem
  • Dye terminator

    as90, as120 on H-stem
  • Complete!