Team:Tokyo Metropolitan/Notebook/Fiber/2010/10/20
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+ | ==2010/10/20 Wednesday (Naoto)== | ||
+ | '''member''' | ||
+ | naoto and watachin | ||
+ | |||
+ | ===Experiment:Sequence=== | ||
+ | '''material''' | ||
+ | |||
+ | *PCR | ||
+ | **bcsD in pSB1C3(No.89~96) | ||
+ | **Big Dye | ||
+ | **primer | ||
+ | **DW | ||
+ | *Ethanol precipitation | ||
+ | **ethanol | ||
+ | **EDTA | ||
+ | |||
+ | '''procedure''' | ||
+ | |||
+ | #mix materials(DNA<50ng) | ||
+ | #PCR | ||
+ | #Add EDTA and Ethanol and put at room temperature (15min) | ||
+ | #centrifuge (8000rpm,30min) | ||
+ | #throw away supernatant | ||
+ | #add ethanol again | ||
+ | #centrifuge (8000rpm,15min) | ||
+ | #put at refrigerator | ||
+ | #throw away supernatant | ||
+ | #add Hi-Di solution | ||
+ | #transfer these sample to plate for sequence | ||
+ | #read sequence | ||
+ | |||
+ | '''result''' | ||
+ | |||
+ | Failure to read sequence |
Revision as of 06:26, 26 October 2010
E.coli Fiber Project Notebook
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2010/10/20 Wednesday (Naoto)
member naoto and watachin
Experiment:Sequence
material
- PCR
- bcsD in pSB1C3(No.89~96)
- Big Dye
- primer
- DW
- Ethanol precipitation
- ethanol
- EDTA
procedure
- mix materials(DNA<50ng)
- PCR
- Add EDTA and Ethanol and put at room temperature (15min)
- centrifuge (8000rpm,30min)
- throw away supernatant
- add ethanol again
- centrifuge (8000rpm,15min)
- put at refrigerator
- throw away supernatant
- add Hi-Di solution
- transfer these sample to plate for sequence
- read sequence
result
Failure to read sequence