Conditions for pyrosequencing are listed below:Forward primer: GAGCAAGGCTGCTAAGTGGAAGACBiotinylated reverse primer: ATGACTTGGAGGCAGCAGACASequencing primer: CCATCCACCCTGAGGAPCR conditions for pyro sequencing:Template: 8 ng genomic DNA in 15uL final reaction volumePrimer: each 0.2uMdNTPs:each 0.125mM;PCR buffer: 1.5ul (10x) AmpliTaq Gold buffer, Mg 1.5mM, Taq Gold Polymerase: 0.02units/ul5 min 95C denature and 10 min 72C final extension; 50 cycles: 20s at 95C, 30s at 62C, 20s at 72C