Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/06
From 2010.igem.org
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===Experiment:electrophoresis=== | ===Experiment:electrophoresis=== | ||
- | '''Member'''<br/> | + | :'''Member'''<br/> |
- | naoto, watachin, bambi75 and NEX | + | :naoto, watachin, bambi75 and NEX |
- | '''Material'''<br/> | + | :'''Material'''<br/> |
- | *PCR production (bcsA and bcsB | + | :*PCR production (bcsA and bcsB) |
- | *1×TAE buffer | + | :*1×TAE buffer |
- | *Agarose gel | + | :*Agarose gel |
- | '''Procedure'''<br/> | + | :'''Procedure'''<br/> |
- | refer to <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol4</a></html> | + | :refer to <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol4</a></html> |
- | '''Result'''<br/> | + | :'''Result'''<br/> |
- | Each bands were not appeared | + | :Each bands were not appeared |
- | === | + | ===Preparation:Making LB plate=== |
- | '''Member'''<br /> | + | :'''Member'''<br /> |
- | naoto | + | :naoto |
- | '''Material''' | + | :'''Material''' |
- | + | ||
- | + | ||
+ | :see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol1</a></html> | ||
- | + | :'''Procedure''' | |
- | ''' | + | |
- | + | ||
- | + | :refer to <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol1</a></html> | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | ''' | + | ===Experiment:PCR=== |
- | + | :'''Member'''<br /> | |
+ | :naoto, watachin, bambi75 and NEX | ||
+ | :'''Material'''<br /> | ||
+ | :*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'''<br /> | |
- | ''' | + | :see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol3</a></html> |
- | + | ||
- | |||
- | |||
- | |||
- | |||
- | + | ===Experiment:Transformation of pSB1C3=== | |
- | + | :'''Member'''<br /> | |
- | + | :Same above | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | ''' | + | |
- | + | ||
+ | :'''Materials'''<br /> | ||
+ | :*pSB1C3(25ng/μl) 1μl | ||
+ | :*Competent cell JM109 50μl | ||
+ | :*LB + Chloramphenicol | ||
+ | :'''Procedure'''<br /> | ||
+ | :#Mix pSB1C3 and competent cell | ||
+ | :#On ice (30min) | ||
+ | :#Heat shock 42℃ 45sec | ||
+ | :#On ice (2min) | ||
+ | :#Inoculate onto LB plates | ||
+ | :#Incubate plates at 37℃ | ||
+ | :'''Consequence'''<br /> | ||
+ | :Failure | ||
<br/> | <br/> |
Revision as of 06:33, 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