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

From 2010.igem.org


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 Thursday(Bambi75)

Experiment:PCR

Member
NEX , Bambi75 and watachin
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 ★.