5.2 Genotyping Salk mutants (PCR)
5.2 Genotyping Salk mutants (PCR) kdorfman Thu, 03/24/2022 - 18:15Check extracts for DNA
- 1% gels
- HW Massruler
Amplify extracted DNA 24 PCR reactions per pair (12 WT + 12 Salk)
| Reagent | WT allele master mix uL | mutant master mix uL | per pair | per class |
|---|---|---|---|---|
| H2O | 243.25 | 243.25 | 486.5 | ~6 mL |
| 10X ex taq buf | 35 | 35 | 70 | ~145 uL |
| dNTP | 28 | 28 | 60 | 720 uL |
| LBP | 17.5 | 17.5 | 210 uL | |
| GSP 1 | 17.5 | ? | ||
| GSP 2 | 17.5 | ? | ||
| Taq 5U/uL | 1.75 uL | 1.75 uL | 3.5 uL | 42 uL |