Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/06
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==2010/09/06 Monday(NEX)== | ==2010/09/06 Monday(NEX)== | ||
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:naoto, watachin, bambi75 and NEX | :naoto, watachin, bambi75 and NEX | ||
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- | + | :refer to <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol1</a></html> | |
===Experiment:PCR=== | ===Experiment:PCR=== | ||
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Latest revision as of 06:34, 27 October 2010
E.coli Fiber Project Notebook
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2010/09/06 Monday(NEX)
Experiment:Electrophoresis
- Member
- naoto, watachin, bambi75 and NEX
- Material
- PCR production (bcsA and bcsB)
- 1×TAE buffer
- Agarose gel
- Procedure
- refer to protocol4
- Result
- Each bands were not appeared
Preparation:Making LB plate
- Member
- naoto
- Material
- see protocol1
- Procedure
- refer to protocol1
Experiment:PCR
- Member
- naoto, watachin, bambi75 and NEX
- Material
- sterilized water 213μl
- Ex taq buffer 30μl
- dNTP 24μl
- Ex taq 3μl
- K12bcsA sense(10μmol/l)5μl
- K12bcsA antisense(10μmol/l)5μl
- K12bcsB sense(10μmol/l)5μl
- K12bcsB antisense(10μmol/l)5μl
- K12bcsC sense(10μmol/l)5μl
- K12bcsC antisense(10μmol/l)5μl
- E.coli K12 strain
- Procedure
- see protocol3
Experiment:Transformation of pSB1C3
- Member
- Same above
- Materials
- pSB1C3(25ng/μl) 1μl
- Competent cell JM109 50μl
- LB + Chloramphenicol
- Procedure
- Mix pSB1C3 and competent cell
- On ice (30min)
- Heat shock 42℃ 45sec
- On ice (2min)
- Inoculate onto LB plates
- Incubate plates at 37℃
- Consequence
- Failure