From 2010.igem.org
E.coli Fiber Project Notebook
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℃
"
