Team:Tokyo Metropolitan/Notebook/Fiber/2010/10/19

From 2010.igem.org


E.coli Fiber Project Notebook

August 2010
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2010/10/19 Tuesday (Naoto)

member
naoto and watachin

Experiment:Colony PCR

material
  • colony of E.coli
    • bcsB:1~96
    • bcsC:97~196
other materials were same as protocol3
procedure
see protocol3

Experiment:Electrophoresis

material
  • PCR production (Colony PCR)
you can see other materials at protocol4
procedure
refer to protocol4
result
We couldn't see bands of bcsB and bcsC...

Experiment:Sequence

material
  • bcsD in pSB1C3(No.89~96)
  • Big Dye
  • primer
  • DW
  • 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
result
Failure to read sequence

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