Table 1 PCR conditions
PCR purpose | PCR conditions (initial denaturation, number of cycles, denaturation, annealing, elongation, and a final extension step) |
|---|---|
Enrichment for microsatellite-containing fragments | 94°C for 2 min, followed by 12 cycles of 94°C for 15 sec, 55°C for 35 sec, 72°C for 90 sec |
Recovery of microsatellite-containing fragments | 35 cycles of 94°C for 15 sec, 55°C for 35 sec, 72°C for 30 sec, and a final extension at 72°C for 5 min |
Preparation of products for cloning | 94°C for 2 min, followed by 15 cycles of 94°C for 15 sec, 55°C for 35 sec, 72°C for 30 sec, and a final extension at 72°C for 5-10 min |
Confirmation of cloned products | 96°C for 2 min, followed by 33 cycles of 94°C for 40 sec, 57°C for 12 sec, 72°C for 30 sec, and a final extension at 72°C for 5 min |
Genotyping with SSRs | 96°C for 2 min, followed by 33 cycles at 94°C for 35 sec, annealing temperature* for 15 sec, 72°C for 30 sec, and a final extension at 72°C for 5 min |