Team:HokkaidoU Japan/Notebook/Length/as90 120

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Notebook
Lab Documents

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-vector 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 on H-stem-vector XhoI_RBS_F Tm:68.2℃ as90_NcoI_R Tm:79.9℃
    as120 on H-stem-vector XhoI_RBS_F Tm:68.2℃ as120_NcoI_R Tm:66.9℃

  • Liquid Culture

    as90, as120 on H-stem

  • Mini-prep

    as90, as120 on H-stem

  • Dye terminator

    as90, as120 on H-stem

  • Complete!

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