Conditions for pyrosequencing are listed below:Biotinylated Forward primer: TCTTTTCAAAACCCCCTTTTCCReverse primer: ACCATAGTTTTGCCTCCTCTGCSequencing primer: TTAACAACAGAGCATT (reverse direction)PCR conditions for pyro sequencing:Template: 5 ng genomic DNA in 15uL final reaction volumePrimer: each 0.2uMdNTPs:each 0.125mM;PCR buffer: 1.5ul (10x) AmpliTaq Gold buffer, Mg 2.5mM, Taq Gold Polymerase: 0.02units/ul5 min 95C denature and 10 min 72C final extension; 50 cycles: 20s at 95C, 30s at 61C, 20s at 72C