PCR reactions containing 40 ng of template DNA, 2 μM each of the four dNTPs, 1 × PCR buffer (10 mM Tris pH 9.0, 50 mM KCl), 2.5 mM of MgCl2, 1 U of Taq polymerase (Bangalore Genei, India) and 20 pmol of primer, were carried out in 25 μl volumes of 0.2 ml microfuge tubes (Dialabs).