Each reaction mixture contained 500 ng of extracted template DNA, 0.4 lM of each primer,200 lM of dNTPs, and 2.5 U of Pro Taq DNA polymerase (Amresco,Solon, Ohio, USA) with a buffer containing 20 mM Tris–HCl, pH 8.0,15 mM MgCl2, 1% Triton X-100, 500 mM KCl, and 0.1% (w/v) gelatin.A negative control reaction (without DNA) was also included for each PCR set to test for possible contamination