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

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Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-GFP-dt on pSB6A1 on ice  30min
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Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-B0040-B0015(on pSB6A1)
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each of 2.5&micro;L DNA to DH5&alpha; 42C    30sec
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    each of 2.5&micro;L DNA to DH5&alpha;
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Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(pSB6A1) pHN1257_up_F Tm:60.1&#8451; & asB0034-R Tm:68.7&#8451;
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fluoresence imageing 488 nm 530 filter
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fluorescence imaging 488 nm 530 filter
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Latest revision as of 15:31, 9 September 2015

Notebook
Lab Documents

140910 asB0034 & B0034 GFP Assay

  • Start

  • Double Transformation

    Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-B0040-B0015(on pSB6A1)
    each of 2.5µL DNA to DH5α

  • Colony PCR

    Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(pSB6A1) pHN1257_up_F Tm:60.1℃ & asB0034-R Tm:68.7℃

  • Preculture

    Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-B0040-B0015(on 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

    fluorescence imaging 488 nm 530 filter

  • Complete!

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