Team:Marburg/Protocols/Protocol13
PCR Standard
GXL-PCR:- prepare PCR mix
| PCR Mix | |
|---|---|
| 5 μL | Primer 1 |
| 5 μL | Primer 2 |
| 24 μL | ddH2O |
| 10 μL | 5x-Puffer |
| 4 μL | dNTPs |
| 2 μL | GXL-Polymerase |
- start cycle (30x)
-
- 98 °C for 10 s
- 55/60 °C (depending on Tm) for 15 s
- 68 °C for 10 s/kb
- end cycle
- store @ 10 °C
Phusion-PCR:
- prepare PCR mix
| PCR Mix | |
|---|---|
| 1 µL | template |
| 1 µL | Primer 1 |
| 1 µL | Primer 2 |
| 35,5 µL | ddH2O |
| 10 µL | 5x-Puffer |
| 1 µL | dNTPs |
| 0,5 µL | Phusion-Polymerase |
- start cycle (30x)
-
- 98 °C for 10 s
- 55/60 °C (depending on Tm) for 15 s
- 68 °C for 10 s/kb
- end cycle
- store @ 10 °C
