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Team:Tokyo Metropolitan/Notebook/Fiber/2010/10/23
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Revision as of 17:08, 25 October 2010
E.coli Fiber Project Notebook
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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
- mix materials(DNA<50ng)
- PCR
- Add EDTA and Ethanol and put at room temperature (15min)
- centrifuge (8000rpm,30min) and throw away supernatant
- add ethanol again
- centrifuge (8000rpm,15min) and throw away supernatant
- add Hi-Di solution
- heat at 95℃
- transfer these sample to plate for sequence
- 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 know other materials,see protocol4
procesure
refer to protocol4
result
From the length of bands
bcsC→No.54 and 64
bcsD→No.75
seem to be correct inserts
