REV and LPDV PCR primers
Version not yet assigned, updated 2026-09-30. https://dustinedwards.info/research/protocols/rev-lpdv-primers
- 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 | Cox et al. 2022 used primers at 200-400 nM final concentration. |
| Reverse primer (10 µM stock) | 10 µM | 1 µL per reaction | 1 µL | Cox et al. 2022 used primers at 200-400 nM final concentration. |
| Eluted DNA | 5 µL per reaction | 5 µL | Cox et al. 2022 used the same master mix in 25 µL reactions with 2 µL of eluted DNA. |
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.
Run the cycling program for the primer set, as in its table below.
Run the product on the 2% agarose gel in TBE beside the NEB 100 bp ladder.
Expected:A band at the set's product size: see where the REV amplicons sit on the genome and, for LPDV, PCR LPDV p31/CA. On the original pages each gel showed a band in the REV-positive lane and none in the negative lane.