Mc4r-Cone Genotyping

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PRIMERS

  • Fwd: CAC TCG GAG CTT CCC TGA CCC AG
  • Rev: GAC CAT GGT TTC CGA CCC ATT
  • Neo Rev: TTC CAA GCC TCT GAG CCC AGA

Make primers at 1uM (Fwd + Rev and Fwd + Neo Rev). PCR primers separately.

PCR PROTOCOL

PCR Step Temperature Time Cycles Notes
1. Initiation/Melting 94 5:00 1
2. Denaturation 94 0:45 40
3. Annealing 66 0:45 40
4. Elongation 72 0:45 40 steps 2-4 to be run in sequence
5. Amplification 72 7:00 1
6. Finish 4 infinite n/a