Basic techniques in Molecular biology
Polymerase chain reaction
| Total volume | 50/μL |
|---|
| DDW | 36 |
| 10×Buffer | 5 |
| dNTP（2.5mM） | 5 |
| FP（10μM） | 1 |
| RP（10μM） | 1 |
| DNA template | 1 |
| EasyPfu DNA Polymerase | 1 |
Thermocycling
The PCRmachine should be set to run the following steps
| Pre-denaturation | 94℃ | 5min |
|---|
| transsexual | 94℃ | 30s | 30circles |
| annealing | Tm-5℃ | 30s |
| extend | 72℃ | 0.5kb/min |
| extend | 72℃ | 5min |
