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