Conditions for pyrosequencing are listed below:Biotinylated forward primer: GCGCTCACCTGCCAGACTReverse primer: TCCCTTTCCTGCGTGACGTSequencing primer (reverse direction): CCGGCGCATGGCTTCPCR conditions for pyro sequencing:Template: 6 ng genomic DNA in 15uL final reaction volumePrimer: each 0.27uMdNTPs:each 0.125mM;PCR buffer: 1.5ul (10x) AmpliTaq Gold buffer, Mg 1.5mM, Taq Gold Polymerase: 0.027units/ul5 min 95C denature and 10 min 72C final extension; 50 cycles: 20s at 95C, 30s at 61C, 20s at 72C