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Team:Tokyo Metropolitan/Notebook/Fiber/2010/08/12
From 2010.igem.org
(Difference between revisions)
Revision as of 09:10, 12 August 2010
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℃
