Team:HokkaidoU Japan/Notebook/asB0034/140910 asB0034&B0032 mRFP Assay

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Notebook
Lab Documents

140910 asB0034 & B0032 mRFP Assay

  • Start

  • Double Transformation

    Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0032-E1010-dt(on pSB6A1)
    each of 2.5µL DNA to DH5α

  • Colony PCR

    R0040-B0032-E1010-B0015(on pSB6A1) EcoRI_XbaI_F Tm:70.9℃ & RBS_SpeI_R Tm:59.2℃

  • Preculture

    Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0032-E1010-B0015(on pSB6A1)
    LB 2 mL for 8hours
    Turbidity was controlled 0.1

  • Culture

    M9ZB 2 mL IPTG 20 µ or not for 9 hours, 11 hours, 13hours

  • Assay

    fluoresence imageing 514 nm 610 filter

  • Complete!

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