Primary Dop Pcr Reaction

To each tube of flow sorted chromosomes (500 sorted chromosomes in 33 ^l H2O):

TAPS2 buffer 5 |l

DOP2 primer (20 |M) 5 |l dNTP mix (2.5 mM each) 4 |l

Polyoxyethelene ether W1 (1%) 2.5 |l

AmpliTaq 0.5 |l

PCR reactions are performed in Biometra thermocyclers. After initial denaturation at 94°C for 10 min, the reaction is as follows: 10 cycles of 94°C for 1.5 min, 30°C for 2.5 min, ramp at 0.23°C s-1 to 72°C, 72°C for 3 min followed by 30 cycles of 94°C for 1 min, 62°C for 1.5 min, 72°C for 3 min, and a final extension step of 72°C for 8 min.

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