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

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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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2010/10/23 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

result

From the length of bands

bcsC→No.54 and 64

bcsD→No.75

seem to be correct inserts



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