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

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E.coli Fiber Project Notebook

August 2010
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2010/10/23 Saturday (Naoto)

member

naoto and watachin

Experiment:Sequence

material

For PCR

  • bcsA (No.9)
  • Big Dye
  • primer
  • DW

For Ethanol precipitation

  • ethanol
  • EDTA

procedure

  1. mix materials(DNA<50ng)
  2. PCR
  3. Add EDTA and Ethanol and put at room temperature　(15min)
  4. centrifuge (8000rpm,30min)
  5. throw away supernatant
  6. add ethanol again
  7. centrifuge (8000rpm,15min)
  8. put at refrigerator
  9. throw away supernatant
  10. add Hi-Di solution
  11. transfer these sample to plate for sequence
  12. read sequence








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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