Primer | RecA |
| Orf-hisG |
|
|
Step | Temperature | Time | Temperature | Time | Cycles |
Initial denaturation | 94˚C | 5 min | 94˚C | 5 min | |
Touchdown denaturation | 94˚C | 30 sec | 94˚C | 30 sec |
|
Touchdown annealing | 67˚C to 57˚C | 1 min 30 sec | 53˚C to 43˚C | 2 min | 20 |
Touchdown extension | 72˚C | 2 min | 72˚C | 2 min |
|
Regular cycle denaturation | 94˚C | 30 sec | 94˚C | 30 sec | |
Regular cycle annealing | 58.5˚C | 1 min 30 sec | 47˚C | 2 min | 20 |
Regular cycle extension | 72˚C | 2 min | 72˚C | 2 min |
|
Final extension | 72˚C | 5 min | 72˚C | 5 min | 1 |