Team:Tokyo Metropolitan/Notebook/Fiber/2010/08/12
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- | {{:Team:Tokyo_Metropolitan/ | + | {{:Team:Tokyo_Metropolitan/Notebook/Fiber}} |
- | ==2010/08/12 Tuesday (Bambi75) | + | ==2010/08/12 Tuesday (Bambi75)== |
===Experiment:PCR=== | ===Experiment:PCR=== | ||
- | ''' | + | :'''Member''' |
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- | + | :NEX , bambi75 and watachin | |
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- | ''' | + | :'''Materials''' |
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- | + | :*sterilized water 106.5μl | |
- | + | :*Ex taq buffer 15μl | |
+ | :*dNTP 12μl | ||
+ | :*Ex taq 1.5μl | ||
+ | :*K12bcsA sense(10μmol/l)2.5μl | ||
+ | :*K12bcsA antisense(10μmol/l)2.5μl | ||
+ | :*K12bcsB sense(10μmol/l)2.5μl | ||
+ | :*K12bcsB antisense(10μmol/l)2.5μl | ||
+ | :*K12bcsC sense(10μmol/l)2.5μl | ||
+ | :*K12bcsC antisense(10μmol/l)2.5μl | ||
+ | :*E.coliK12strain | ||
- | + | :'''Equipment''' | |
+ | :*thermal cycler | ||
+ | :*vortex mixer | ||
+ | :*PCR-tubes | ||
+ | :*pipette tip | ||
- | + | :'''Procedure''' | |
- | + | :1.mix materials : sterilized water , Ex taq buffer , dNTP and Ex taq and divide 1) into 3 tips. | |
+ | :2.mix K12bcsAsense , K12bcsAantisense and 2)<br/> | ||
+ | :mix K12bcsBsense , K12bcsBantisense and 2)<br/> | ||
+ | :mix K12bcsCsense , K12bcsCantisense and 2)<br/> | ||
+ | :3.dip E.coliK12strain into 3). | ||
+ | :4.setting tips and elongation in thermal cycler | ||
+ | :*initialization 95℃ 3min | ||
+ | :*denaturation 96℃1min | ||
+ | :*annealing 55℃ 1min | ||
+ | :*elongation 72℃ 5min (30 cycles from 95℃ to 72℃) | ||
+ | :*reaction stop 10℃ | ||
- | + | <br/> | |
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Latest revision as of 21:24, 26 October 2010
E.coli Fiber Project Notebook
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2010/08/12 Tuesday (Bambi75)
Experiment:PCR
- Member
- NEX , bambi75 and watachin
- Materials
- sterilized water 106.5μl
- Ex taq buffer 15μl
- dNTP 12μl
- Ex taq 1.5μl
- K12bcsA sense(10μmol/l)2.5μl
- K12bcsA antisense(10μmol/l)2.5μl
- K12bcsB sense(10μmol/l)2.5μl
- K12bcsB antisense(10μmol/l)2.5μl
- K12bcsC sense(10μmol/l)2.5μl
- K12bcsC antisense(10μmol/l)2.5μl
- E.coliK12strain
- Equipment
- thermal cycler
- vortex mixer
- PCR-tubes
- pipette tip
- Procedure
- 1.mix materials : sterilized water , Ex taq buffer , dNTP and Ex taq and divide 1) into 3 tips.
- 2.mix K12bcsAsense , K12bcsAantisense and 2)
- mix K12bcsBsense , K12bcsBantisense and 2)
- mix K12bcsCsense , K12bcsCantisense and 2)
- 3.dip E.coliK12strain into 3).
- 4.setting tips and elongation in thermal cycler
- initialization 95℃ 3min
- denaturation 96℃1min
- annealing 55℃ 1min
- elongation 72℃ 5min (30 cycles from 95℃ to 72℃)
- reaction stop 10℃