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For PCR amplification of pan-avian GAPDH.
Forward primer: 5’•GTGGTGCTAAGCGTGTTATCATC•3’
Reverse primer: 5’•GGCAGCACCTCTGCCATC•3’
Materials
- NEB OneTaq® Hot Start 2X Master Mix
- NEB 100 bp ladder
- 2% agarose gel in TBE
Extension temperature is dependent on polymerase
| Step | Temperature (°C) | Time |
| initial denaturation | 95 | 10 min. |
| 40 cycles | 95 | 30 sec. |
| 58 | 60 sec. | |
| 68 | 30 sec. | |
| extension | 68 | 5 min. |
| hold | 10 | ∞ |
