which was developed by Hirano and Arie[20]. Each amplification
was performed in a 25mL reaction volume containing 20 mM
Tris–HCl (pH 8.4), 50 mM KCl, 2 mM MgCl2,800mM dNTP
(dATP:dTTP:dCTP:dGTP in a 1:1:1:1 ratio), 0.25mM of primers, 1 U
of PlatinumTaq polymerase (Invitrogen, Life Technologies) and
15 ng of template DNA