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Team:Tokyo Metropolitan/Notebook/Fiber/2010/10/19
From 2010.igem.org
(Difference between revisions)
| Line 30: | Line 30: | ||
We couldn't see bands of bcsB and bcsC... | 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''' | ||
| + | |||
| + | #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 | ||
<br/> | <br/> | ||
Revision as of 18:01, 26 October 2010
E.coli Fiber Project Notebook
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Contents |
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
- 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
