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Team:Tokyo Metropolitan/Notebook/Fiber/2010/08/12
From 2010.igem.org
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{{:Team:Tokyo_Metropolitan/Header}} | {{:Team:Tokyo_Metropolitan/Header}} | ||
| - | === | + | ==2010/08/12 Tuesday (Bambi75) |
| + | ===Experiment:PCR=== | ||
| + | |||
| + | '''member''' | ||
NEX , bambi75 and watachin | NEX , bambi75 and watachin | ||
| - | + | '''Materials''' | |
*sterilized water 106.5μl | *sterilized water 106.5μl | ||
*Ex taq buffer 15μl | *Ex taq buffer 15μl | ||
| Line 17: | Line 20: | ||
*E.coliK12strain | *E.coliK12strain | ||
| - | + | '''Equipment''' | |
*thermal cycler | *thermal cycler | ||
*vortex mixer | *vortex mixer | ||
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*pipet tip | *pipet tip | ||
| - | + | '''Procedure''' | |
1)mix materials : sterilized water , Ex taq buffer , dNTP and Ex taq. | 1)mix materials : sterilized water , Ex taq buffer , dNTP and Ex taq. | ||
Revision as of 16:25, 24 October 2010
==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
- pipet tip
Procedure 1)mix materials : sterilized water , Ex taq buffer , dNTP and Ex taq.
2)divide 1) into 3 tips.
3)mix K12bcsAsense , K12bcsAantisense and 2).
mix K12bcsBsense , K12bcsBantisense and 2).
mix K12bcsCsense , K12bcsCantisense and 2).
4)dip E.coliK12strain into 3).
5)setting tips and elongation in thermal cycler
- initialization 95℃ 3min
- denaturation 96℃1min
- annealing 55℃ 5min
- elongation 72℃ 1min (30 cycles from 95℃ to 72℃)
- reaction stop 10℃
