Team:HokkaidoU Japan/Notebook/asB0034/asB0034-construction

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Ptrc-stem-asB0034-stem(on pHN1257) pHN1257_up_F Tm:60.1$&#8451; & asB0034-R Tm:68.7&#8451;
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Ptrc-stem-asB0034-stem(on pHN1257) pHN1257_up_F Tm:60.1&#8451; & asB0034-R Tm:68.7&#8451;
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Revision as of 00:59, 18 October 2014

Notebook
Lab Documents

asB0034 Construction

  • Start

  • Primer Annealing

    asB0034_F Tm:68.7℃ asB0034_R Tm:68.7℃

  • Digestion

    Cut pHN1257 with NcoI, XhoI(using 10×cut smart)

  • Gel extraction

    digested pHN1257

  • Ethanol Precipetation

    digested pHN1257

  • Ligation

    Ligate pHN1257 with asB0034

  • Transformation

    Ptrc-stem-asB0034-stem on pHN1257 5.0µL DNA to DH5α

  • Colony PCR

    Ptrc-stem-asB0034-stem(on pHN1257) pHN1257_up_F Tm:60.1℃ & asB0034-R Tm:68.7℃

  • Culture

    Ptrc-stem-asB0034-stem(on pHN1257)

  • mini-Prep

    Ptrc-stem-asB0034-stem(on pHN1257)

  • Double Transformation

    Ptrc-stem-asB0034-stem(on pHN1257)&R0040-B0034-E1010-B0015(on pSB6A1) each of 2.5µL DNA to DH5α

  • Complete!

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