Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/01

From 2010.igem.org


E.coli Fiber Project Notebook

August 2010
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2010/09/01 Wednesday(Naoto)

Experiment:Digestion of bcsA,B,C and D

Member
naoto
Material
  • bcsA,B,C,D
  • 10×Mbuffer
  • BSA
  • XbaI
  • SpeI
Procedure
  1. add "material" to PCR tubes(show below)
  2. incubation at (37℃,7h)
composition
bcsAbcsBbcsCbcsD
solution of bcs(μl) 20202020
10×Mbuffer(μl) 2 2 2 2
BSA(μl) 2 2
XbaI(μl) 0.8 0.8
SpeI(ul) 0.8 0.8

Preparation:Subculture ofA.xylinum

Member
naoto
Material
A.xylinum JCM7664
Procedure
transfer A.xylinum JCM7664 to new culture(OWW and Broth 8/25 made)

Experiment:PCR

Member
naoto
Material
  • E.coli K12 strain
  • 2×PCR buffer 25μl×2
  • 2mM dNTP 10μl×2
  • 10μM primer(sense)bcsA,B 2.5μl each
  • 10μM primer(antisense)bcsA,B 2.5μl each
  • milli-Q water 9μl×2
  • KOD FX 0.5μl×2
Procedure
follow protocol3

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