Ferment It Yourself

Vitamin A	Vitamin B2	Vitamin B12
Phytase	Riboswitch	Differentiation on E. coli
Differentiation on S. cerevisiae	Manufacturing	Idli and Micro-organisms

PCR protocol (Phusion)

Thaw PCR Master Mix on ice for 10min.
add 2µL of each primers (at 10 µM) in a PCR tube.
add 1µL of the DNA (at 1 ng/µL) you want to amplify in the same tube.
add 25µL of Life Technologies Phusion High-Fidelity PCR Master Mix with HF Buffer.
add 20µL of DNase/RNase-free water to the tube.
Thermocycling conditions
- initial denaturation 98°C for 30 seconds
- 35 cycles
  - 98°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

Team:Paris Bettencourt/Protocols/PCR