PCR protocol (Phusion)
- Thaw PCR mastermix on ice for 10min.
- add 2µL of each primers in a PCR tube.
- add 1uL of the DNA you want to amplify in the same tube.
- add 25uL of PCR master mix (Phusion).
- add 20uL of water to the tube.
- Thermocycling conditions
- initial denaturation 98°C for 30 seconds
- 35 cycles
- 94°C for 30 seconds
- 57°C for 30 seconds
- 72°C 30 seconds/kb
- final extension of 72°C for 10 minutes
- infinite hold 12°C
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