Difference between revisions of "GDF15 Genotyping"
From Bridges Lab Protocols
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Latest revision as of 14:51, 8 June 2020
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 |
