Table 6.
PCR cycling conditions for KOD HOT Start DNA Polymerase for HA cassette amplification
| Steps | Temperature | Time | Cycles |
|---|---|---|---|
| Polymerase activation | 95°C | 2 min | 1 |
| 1ststep (Primers annealing to the HA cassette and initial amplification) | |||
| Denaturation | 95°C | 20 s | 5 |
| Annealing | (According to primers designed by the user) °C | 20 s | |
| Extension | 70°C | 45 s |
|
| 2ndstep (Final amplification) | |||
| Denaturation | 95°C | 20 s | 30 |
| Annealing | (Average of full-length Frw and Rev primers Tm) °C | 20 s | |
| Extension | 70°C | 45 s | |
| Final extension | 70°C | 3 min | 1 |
| Hold | 4°C | ꝏ | |
See also Figure 2.