Team:Tokyo Metropolitan/Notebook/Fiber/2010/10/23

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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2010/10/23 Saturday (Naoto)

member
naoto and watachin

Experiment:Sequence

  • bcsA (No.9)
  • Big Dye
  • primer
  • DW
  • ethanol
  • EDTA
  • Hi-Di solution
procedure
  1. mix materials(DNA<50ng)
  2. PCR
  3. Add EDTA and Ethanol and put at room temperature　(15min)
  4. centrifuge (8000rpm,30min) and throw away supernatant
  5. add ethanol again
  6. centrifuge (8000rpm,15min) and throw away supernatant
  7. add Hi-Di solution
  8. heat at 95℃
  9. transfer these sample to plate for sequence
  10. read sequence
result
bcsA wasn't inserted into pSB1C3...

Experiment:Colony PCR

material
  • colony of E.coli
    • bcsB:1~32
    • bcsC:33~64
    • bcsD:65~96
other materials were same as protocol3
procedure
see protocol3

Experiment:Electrophoresis

material
  • PCR production (Colony PCR)
you can see other materials at protocol4
procesure
refer to protocol4
result
From the length of bands
bcsC→No.54 and 64
bcsD→No.75
seem to be correct inserts



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