Table 2 Thermal cycling parameters used in SWPOP-PCR method
From: Stepwise partially overlapping primer-based PCR for genome walking
Round of PCR | Stage | Thermal condition | Cycle number |
|---|---|---|---|
Primary | 94 °C 2 min | 1 | |
1 | 94 °C 30 s, 65 °C 1 min, 72 °C 2 min | 5 | |
2 | 94 °C 30 s, 25 °C 1 min, 72 °C 2 min | 1 | |
3 | 94 °C 30 s, 65 °C 1 min, 72 °C 2 min | 25 | |
72 °C 5 min | 1 | ||
1 μL of the product was directly used as plate in the secondary round of PCR | |||
Secondary | 94 °C 2 min | 1 | |
1 | 94 °C 30 s, 65 °C 1 min, 72 °C 2 min | 5 | |
2 | 94 °C 30 s, 50 °C 1 min, 72 °C 2 min | 1 | |
3 | 94 °C 30 s, 65 °C 1 min, 72 °C 2 min | 25 | |
72 °C 5 min | 1 | ||
1 μL of the product was directly used as plate in the tertiary round of PCR | |||
Tertiary | Reaction profile of tertiary PCR is identical to that of secondary PCR | ||