Difference between revisions of "Ppp1r3c Genotyping"
From Bridges Lab Protocols
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Latest revision as of 18:17, 5 June 2020
PRIMERS
- Fwd: TGCTCTCCTTTTCTCCACGT
- Rev: CCGGACCTGAACCTTCTTCT
- Neo Rev: GCAGCGCATCGCCTTCTATC
Make primers at 1uM (Fwd + Rev and Fwd + Neo Rev}. PCR primers separately.
PCR PROTOCOL
| PCR Step | Temperature | Time | Cycles | Notes |
|---|---|---|---|---|
| 1. Initiation/Melting | 95 | 3:00 | 1 | |
| 2. Denaturation | 95 | 0:30 | 35 | |
| 3. Annealing | 60 | 1:00 | 35 | |
| 4. Elongation | 72 | 1:00 | 35 | steps 2-4 to be run in sequence |
| 5. Amplification | 72 | 5:00 | 1 | |
| 6. Finish | 4 | infinite | n/a |
