Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/06

From 2010.igem.org


E.coli Fiber Project Notebook

August 2010
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September 2010
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2010/09/06 Monday(NEX)

Experiment:Electrophoresis

Member
naoto, watachin, bambi75 and NEX
Material
  • PCR production (bcsA and bcsB)
  • 1×TAE buffer
  • Agarose gel
Procedure
refer to protocol4
Result
Each bands were not appeared

Preparation:Making LB plate

Member
naoto
Material
see protocol1
Procedure
refer to protocol1

Experiment:PCR

Member
naoto, watachin, bambi75 and NEX
Material
  • sterilized water 213μl
  • Ex taq buffer 30μl
  • dNTP 24μl
  • Ex taq 3μl
  • K12bcsA sense(10μmol/l)5μl
  • K12bcsA antisense(10μmol/l)5μl
  • K12bcsB sense(10μmol/l)5μl
  • K12bcsB antisense(10μmol/l)5μl
  • K12bcsC sense(10μmol/l)5μl
  • K12bcsC antisense(10μmol/l)5μl
  • E.coli K12 strain
Procedure
see protocol3


Experiment:Transformation of pSB1C3

Member
Same above
Materials
  • pSB1C3(25ng/μl) 1μl
  • Competent cell JM109 50μl
  • LB + Chloramphenicol
Procedure
  1. Mix pSB1C3 and competent cell
  2. On ice (30min)
  3. Heat shock 42℃ 45sec
  4. On ice (2min)
  5. Inoculate onto LB plates
  6. Incubate plates at 37℃
Consequence
Failure

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