Table 2 PCR conditions for amplification... | American Society of Hematology

Table 2

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

Each reaction was performed by the 3-step PCR method. The PCR was started from the initial denaturation, followed by cycling reactions and incubation.

†

Alternative condition of PCR4 was used for direct sequencing of the shorter product derived from the B^m allele.

View Large

This Feature Is Available To Subscribers Only