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Team:Chiba/protocol/PCR
From 2008.igem.org
PCR
- Resuspend primer in Nuculease free water to 100 µM
- PCR mix
- DNA template 1µL
- Fwd primer 10µL (final con. 10 pM)
- Rev primer 10µL
- 10x thermo pol buffer 10µL
- dNTP mix 10µL
- DNA pol. 1µL
- dH20 58µL
----------------------------
100µL
- PCR cycle
Start: 94 °C for 5 min. (melt)
cycle: melt: 1 min.
anneal : 30 sec.
cycle end: extension:72 °C for 3.5 min.
25 cycles
72 °C for 10 min
store: keep at 6 °C forever
