"
Protocol: PCR
- Get the following excel: File:Team-EPF-Lausanne Protocol PCR.xls
- Fill in the pink fields with the appropriate values.
- The annealing temperature is generally the lowest Tm. For long primers, use lowest Tm-5
- The extension time is 15-30s per 1kb
- Prepare the master mix by adding the elements in the following order:
- H20
- 5x HF Buffer
- dNTP
- Polymerase (keep it cold!)
- Put 50ul of the master mix in a PCR tube for each different PCR
- Add the primers and the DNA
- Start the PCR!
