IGEM:Peking/2007/Count-Conjugation-Notebook/2007-7-25

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(New page: ==R0040<-crRNA== *Ligation transformants do not grow. *Re-ligation tonight, and overnight, plan to ligate under 16℃ tomorrow morning. *In case it is the XbaI/PstI digestion of the PCR pr...)
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Revision as of 11:20, 25 July 2007

R0040<-crRNA

  • Ligation transformants do not grow.
  • Re-ligation tonight, and overnight, plan to ligate under 16℃ tomorrow morning.
  • In case it is the XbaI/PstI digestion of the PCR product ends that failed. We will rePCR tonight, see below.

R0010<-J23066

  • Cut J23066 with XbaI/PstI and R0040 with SpeI/PstI.
  • Electrophoresis and gel extraction of J23066.
  • Ligation over night.

rePCR crRNA

  • By Yang Yifan
  • J23078 primer1 and primer2 stored as ~50uM
  • PCR 5 cycle. total volume: 50ul
  • Primer final concentration 0.5uM. Last time, we used 1uM, a little more according to electrophoresis yesterday.
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