Mc4r-Cone Genotyping
From Bridges Lab Protocols
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 |
