PCR conditions for amplification of genomic DNA
PCR* . | Initial denaturation . | Cycles; denaturation . | Annealing . | Extension . | Incubation . |
---|---|---|---|---|---|
Region −8.8 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 60°C for 15 s | 68°C for 4 min | 68°C for 7 min |
Region +5.8 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 60°C for 15 s | 68°C for 3 min | 68°C for 7 min |
Region +11.5 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 60°C for 15 s | 68°C for 4 min | 68°C for 7 min |
PCR1-PCR3 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 60°C for 15 s | 68°C for 16 min | 68°C for 7 min |
PCR4 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 66°C for 15 s | 68°C for 8 min | 68°C for 7 min |
PCR4† | 94°C for 3 min | 35 cycles; 98°C for 10 s | 66°C for 15 s | 68°C for 1 min | 68°C for 7 min |
PCR5 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 66°C for 15 s | 68°C for 8 min | 68°C for 7 min |
PCR6 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 66°C for 15 s | 68°C for 6 min | 68°C for 7 min |
PCR7 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 66°C for 15 s | 68°C for 10 min | 68°C for 7 min |
PCR8 and PCR9 | 94°C for 3 min | 30 cycles; 98°C for 1 min | 55°C for 48 s | 72°C for 3.4 min | 72°C for 10 min |
PCR* . | Initial denaturation . | Cycles; denaturation . | Annealing . | Extension . | Incubation . |
---|---|---|---|---|---|
Region −8.8 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 60°C for 15 s | 68°C for 4 min | 68°C for 7 min |
Region +5.8 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 60°C for 15 s | 68°C for 3 min | 68°C for 7 min |
Region +11.5 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 60°C for 15 s | 68°C for 4 min | 68°C for 7 min |
PCR1-PCR3 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 60°C for 15 s | 68°C for 16 min | 68°C for 7 min |
PCR4 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 66°C for 15 s | 68°C for 8 min | 68°C for 7 min |
PCR4† | 94°C for 3 min | 35 cycles; 98°C for 10 s | 66°C for 15 s | 68°C for 1 min | 68°C for 7 min |
PCR5 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 66°C for 15 s | 68°C for 8 min | 68°C for 7 min |
PCR6 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 66°C for 15 s | 68°C for 6 min | 68°C for 7 min |
PCR7 | 94°C for 3 min | 35 cycles; 98°C for 10 s | 66°C for 15 s | 68°C for 10 min | 68°C for 7 min |
PCR8 and PCR9 | 94°C for 3 min | 30 cycles; 98°C for 1 min | 55°C for 48 s | 72°C for 3.4 min | 72°C for 10 min |