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Team:Tokyo Metropolitan/Notebook/Fiber/2010/08/12
From 2010.igem.org
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:1.mix materials : sterilized water , Ex taq buffer , dNTP and Ex taq and divide 1) into 3 tips. | :1.mix materials : sterilized water , Ex taq buffer , dNTP and Ex taq and divide 1) into 3 tips. | ||
| - | :2. | + | :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). | :3.dip E.coliK12strain into 3). | ||
:4.setting tips and elongation in thermal cycler | :4.setting tips and elongation in thermal cycler | ||
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℃
