Team:HokkaidoU Japan/Notebook/asB0034/140910 asB0034&B0034 GFP Assay

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Notebook
Lab Documents

140910 asB0034 & B0034 GFP Assay

  • Start

  • Transformation

    Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-GFP-dt on pSB6A1 on ice 30min each of 2.5µL DNA to DH5α 42℃ 30sec on ice 2min

  • Colony PCR

    Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1) pHN1257_up_F Tm:60.1℃ asB0034-R Tm:68.7℃

  • Preculture

    Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-GFP-dT(pSB6A1) LB 2 mL for 9hours Turbidity was controlled 0.1

  • Culture

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

  • Assay

    fluoresence imageing 488 nm 530 filter

  • Complete!

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