PCR for genotyping
Material
- Genomic DNA
See Genomic DNA extraction from mouse tail by NaOH or by proteinase K. - 10 uM primer
See Genotyping primer. - DMSO (Invitrogen D8418)
- 25 mM dNTP mix (GenScript C01581, -20C)
- 10 mg/ml BSA (NEB B9001, -20C)
- Taq (NEB, -20C)
Composition
| Item | Vol (ul) |
|---|---|
| gDNA | 1 |
| x10 PCR buf | 2 |
| DMSO | 2 |
| dNTP | 0.32 |
| Primer | 0.8 each |
| BSA | 0.16 |
| Taq | 0.1 |
| DW | to 20 |
PCR
| Temp | Time | Cycle |
|---|---|---|
| 95C | 3 min | |
| 95C 50-65C 65C |
30 sec 30 sec 1 min |
x40 |
| 65C 10C |
7 min ∞ |