Team:Tokyo Metropolitan/Notebook/Fiber/2010/10/23
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==2010/10/23 Saturday (Naoto)== | ==2010/10/23 Saturday (Naoto)== | ||
- | '''member''' | + | :'''member''' |
- | naoto and watachin | + | :naoto and watachin |
===Experiment:Sequence=== | ===Experiment:Sequence=== | ||
- | |||
- | + | :*bcsA (No.9) | |
- | *bcsA (No.9) | + | :*Big Dye |
- | *Big Dye | + | :*primer |
- | *primer | + | :*DW |
- | *DW | + | :*ethanol |
- | + | :*EDTA | |
- | *ethanol | + | :*Hi-Di solution |
- | *EDTA | + | |
- | '''procedure''' | + | :'''procedure''' |
- | #mix materials(DNA<50ng) | + | :#mix materials(DNA<50ng) |
- | #PCR | + | :#PCR |
- | #Add EDTA and Ethanol and put at room temperature (15min) | + | :#Add EDTA and Ethanol and put at room temperature (15min) |
- | #centrifuge (8000rpm,30min) | + | :#centrifuge (8000rpm,30min) and throw away supernatant |
- | + | :#add ethanol again | |
- | #add ethanol again | + | :#centrifuge (8000rpm,15min) and throw away supernatant |
- | #centrifuge (8000rpm,15min) | + | :#add Hi-Di solution |
- | + | :#heat at 95℃ | |
- | + | :#transfer these sample to plate for sequence | |
- | #add Hi-Di solution | + | :#read sequence |
- | #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 <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol3</a></html> | ||
+ | :'''procedure''' | ||
+ | :see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol3</a></html> | ||
+ | ===Experiment:Electrophoresis=== | ||
+ | :'''material''' | ||
+ | :*PCR production (Colony PCR) | ||
+ | :you can see other materials at <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80electrophoresis">protocol4</a></html> | ||
+ | :'''procesure''' | ||
+ | :refer to <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80electrophoresis">protocol4</a></html> | ||
+ | :'''result''' | ||
+ | :From the length of bands | ||
+ | :bcsC→No.54 and 64 | ||
- | + | :bcsD→No.75 | |
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- | + | :seem to be correct inserts | |
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Latest revision as of 18:07, 27 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
- colony of E.coli
- other materials were same as protocol3
- procedure
- see protocol3
Experiment:Electrophoresis
- material
- PCR production (Colony PCR)
- you can see other materials at protocol4
- procesure
- refer to protocol4
- result
- From the length of bands
- bcsC→No.54 and 64
- bcsD→No.75
- seem to be correct inserts