From: DAR-PCR: a new tool for efficient retrieval of unknown flanking genomic DNA
Round of PCR | Stage | Thermal conditions | Cycle number |
---|---|---|---|
Primary | 94 °C, 2 min | ||
1 | 94 °C 30 s, 60 °C 30 s, 72 °C 2 min | 5 | |
2 | 94 °C 30 s, 25 °C 30 s, 72 °C 2 min | 1 | |
3 | 94 °C 30 s, 55 °C 30 s, 72 °C 2 min | 15 | |
4 | 94 °C 30 s, 65 °C 30 s, 72 °C 2 min | 25 | |
72 °C 5 min | |||
1 µL of the product was directly used as template for secondary PCR | |||
Secondary | 94 °C, 2 min | ||
1 | 94 °C 30 s, 60 °C 30 s, 72 °C 2 min | 35 | |
72 °C 5 min |