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Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/06
From 2010.igem.org
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Follow to protocol3 Direct PCR | Follow to protocol3 Direct PCR | ||
| - | ===Experiment=== | + | ===Experiment:Transformation of pSB1C3=== |
'''Member'''<br /> | '''Member'''<br /> | ||
Same above | Same above | ||
'''Materials'''<br /> | '''Materials'''<br /> | ||
| - | * | + | *pSB1C3(25ng/μl) 1μl |
*Competent cell JM109 50μl | *Competent cell JM109 50μl | ||
*LB + chloramphenicol | *LB + chloramphenicol | ||
'''Procedure'''<br /> | '''Procedure'''<br /> | ||
| - | #Mix | + | #Mix pSB1C3 and competent cell |
#On ice (30min) | #On ice (30min) | ||
#Heat shock 42℃ 45sec | #Heat shock 42℃ 45sec | ||
Revision as of 05:52, 8 September 2010
Contents |
2010/09/06 Monday(NEX)
Experiment:subculture of E.coli
Member
naoto, watachin, bambi75 and NEX
Material
- E.coli K12
Procedure
- Pick up a culture of E.coli K12 and streak it to new culture(LB plate)
Experiment:electrophoresis
Member
Same above
Material
- PCR production (bcsA and bcsB from E.coli)
- 1×TAE buffer
- Agarose gel
Procedure
Follow to protocol4
Consequence
Each bands were not appeared
Experiment:Making LB plate
Member
naoto
Material
- Distilled water 200ml
- LB Broth 4g
Experiment:Direct PCR
Member
naoto, watachin, bambi75 and NEX
Materials
- 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
Follow to protocol3 Direct PCR
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 these onto LB plates
- Incubate plates at 37℃
Consequence
Failure
