Team:HokkaidoU Japan/Notebook/H Stem/Downstream Correction

From 2014.igem.org

Notebook
Lab Documents

Stem-Downstream Correction

  • Start

  • Sequencing

    R0010-stem-as-stem-B0015(on pSB1A3) found stem-dn not inserted!

  • PCR

    R0010-stem-as-B0015(on pSB1A3) with NheI_dT_F Tm:67.9℃ & XhoI_RBS_R Tm:68.2℃

  • Digestion

    XhoI-B0015-1A3-R0010-stem-as-XhoI with XhoI

  • Primer Annealing

    stem_downstream_F Tm:℃ & stem_downstream_R

  • Ligation

    XhoI-B0015-1A3-R0010-stem-as-XhoI & stem-dn

  • Transformation

    R0010-stem-as-stem-B0015(on pSB1A3)

  • Colony PCR

    R0010-stem-as-stem-B0015(on pSB1A3) with NcoI_mRFP_F Tm:76.0℃ & 100_up_R Tm:62.6℃

  • Complete!

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