Team:Tokyo Metropolitan/Notebook/Fiber/2010/08/26

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E.coli Fiber Project Notebook

August 2010
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September 2010
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October 2010
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2010/08/26(Bambi75)

Experiment:PCR

member same above

Materials

  • 2×PCR buffer 25×4μl
  • 2mM dNTP 10×4μl
  • 10mM primer(sense)bcsA,B,C and D 2.5μl each
  • 10mM primer(antisense)bcsA,B,C and D 2.5μl each
  • template DNA a little
  • Q water 9×4μl
  • KOD FX 0.5×4μl

Procedure ①mix all materials for 4 tubes.

②elongation

  • bcsA,bcsB and bcsC
    • 94℃ 2min
    • 98℃ 10sec☆
    • 55℃ 30sec
    • 68℃ 4min★
    • 68℃ 7min
    • 10℃　∞
  • bcsD
    • 94℃ 2min
    • 98℃ 10sec☆
    • 55℃ 30sec
    • 68℃ 1min★
    • 68℃ 7min
    • 10℃　∞

※30cycle ☆ to ★.

Experiment:PCR

member

NEX , Bambi75 and watachin

Materials

  • sterilized water 71μl
  • Ex taq buffer 10μl
  • dNTP mix 8μl
  • Ex taq 1μl
  • primer(bcsC sense/antisense) 5μl each

Procedure ①mix materials

②divide ① into 2 tubes.

③elongation

  • 95℃ 3min
  • 96℃ 1min☆
  • 55℃ 7min
  • 72℃ 1min★
  • 10℃ ∞
※30 cycle ☆to★.
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