Team:HokkaidoU Japan/Notebook/September10

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=3 piece ligation つづき=
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=3 piece ligation, continued=
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* 前日にPCRしたHSPとGFP全量を電気泳動にかけたが，くっきりしたバンドがみられなかった
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* PCRでDNAが十分に増幅されなかったものと考え，cycle数を40回に増やしてもう一度PCRからやった
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* Yesterday after purification by [[Team:HokkaidoU_Japan/Protocols|gel extraction]] electrophoresis showed very weak bands
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* これを同様に電気泳動したが，やはり増幅がうまくいかなかった
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* This time increased PCR cycles to 40
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* Templateが悪いと考え，PCRで増やしたこれらのパーツを鋳型に，Extension 90 sec, cycle数30回でもう一度PCRを行った
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* Everything else was same as last time
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* But again it was a failure
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* Thought that template went bad，so used PCR Product as template
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* Extension was 90 sec, did 30 cycles

Latest revision as of 09:24, 2 October 2010

Notebook

3 piece ligation, continued

  • Yesterday after purification by gel extraction electrophoresis showed very weak bands
  • This time increased PCR cycles to 40
  • Everything else was same as last time
  • But again it was a failure
  • Thought that template went bad，so used PCR Product as template
  • Extension was 90 sec, did 30 cycles
Retrieved from "http://2010.igem.org/Team:HokkaidoU_Japan/Notebook/September10"