IGEM:MIT/2007/Protocols

Protocols

PCR

Platinum PCR Supermix

1. Set Reaction Tubes/Plates on Ice
2. Add the following components in a reaction vessel

• • 45 µl Platinum PCR SuperMix
  • Primers (200 nM final concentration per primer is recommended)
  • Template DNA solution

Total volume should be between .5-20 µl.

1. Cap reaction vessel and load into a thermal cycler at 94°C
2. Incubate tubes in a thermal cycler at 94°C for 30 s to 2 min
3. Perform 25-35 cycles of PCR amplification
   1. Denature 94° for 15-30 s
   2. Anneal 55°C for 15-30 s
   3. Extend 72°C for 1 min per kb