GDF15 Genotyping

PRIMERS

  • Fwd1: 5' GAT TCC CGC CCG AAT TAG C 3'
  • Fwd2: 5' CCG AAT TAG CCT GGT CAC CC 3'
  • Rev: 5’ ATC CGT CCT ACT CTG GCT AAG 3'

Make primers at 0.4uM (Fwd1 + Rev and Fwd2 + Rev). PCR primers separately.

PCR PROTOCOL

PCR Step Temperature Time Cycles Notes
1. Initiation/Melting 95 3:00 1
2. Denaturation 95 0:30 38
3. Annealing 60 0:40 38
4. Elongation 72 1:00 38 steps 2-4 to be run in sequence
5. Amplification 72 5:00 1
6. Finish 4 infinite n/a