Pan-avian GAPDH PCR
Version not yet assigned, updated 2026-09-30. https://dustinedwards.info/research/protocols/pan-avian-gapdh
- Updated
- 2026-09-30
Reagents
| Reagent | Stock | Amount | For 1 reaction | Notes |
|---|---|---|---|---|
| Nuclease-free water | 5.5 µL per reaction | 5.5 µL | ||
| One*Taq* Hot Start 2X Master Mix (New England Biolabs) | 12.5 µL per reaction | 12.5 µL | NEB OneTaq® Hot Start 2X Master Mix | |
| Forward primer (10 µM stock) | 10 µM | 1 µL per reaction | 1 µL | |
| Reverse primer (10 µM stock) | 10 µM | 1 µL per reaction | 1 µL | |
| Eluted DNA | 5 µL per reaction | 5 µL |
Equipment
NEB 100 bp ladder; 2% agarose gel in TBE
Reaction mix
Set up each 25 µL reaction as in the table: 5.5 µL of Nuclease-free water, 12.5 µL of OneTaq Hot Start 2X Master Mix (New England Biolabs), 1 µL of Forward primer (10 µM stock), 1 µL of Reverse primer (10 µM stock) and 5 µL of Eluted DNA.
PCR conditions
Run the cycling program in the table above.
Run the product on the 2% agarose gel in TBE beside the NEB 100 bp ladder and look for the band at 534 bp.
Expected:A band at 534 bp. On the original version of this page, the positive lane (DNA from DF-1 cells) showed the product at 534 bp and the negative lane had no band.