A total of 2.5 ml of the purified genomic DNA was used as a template for the PCR amplifications carried out in a total volume of 50 mL containing 0.5 mmol/L of O-150 primer (59-GAT-TYTTCCGRCGAANARCGC-39) and 0.5 mmol/L of biotinylated O-150 primer (59-B-GCNRTRTAAATNTGNAAATTC-39, where B = biotin; N = A, G, C, or T; Y = C or T; and R = A or G). Reaction mixtures also contained 60 mM Tris-HCl, (pH 9.0), 15 mM (NH4) 2 SO4, 2 mM MgCl 2 , 0.2 mM each of dATP, dCTP, dGTP and dTTP, and 2.5 units of Taq polymerase (Invitrogen). Cycling conditions consisted of five cycles of one minute at 94uC, two minutes at 37uC, and 30 seconds at 72uC, followed by 35 cycles of 30 seconds each at 94uC , 37uC , and 72uC. The reaction was completed by incubating at 72uC for six minutes.