The mixturewas run
in a thermal cycler (Applied Biosystems, model 2720) with the
following PCR program: 95 C initial denaturation for 5 min; followed
by 35 cycles of denaturation at 95 C for 20 s, primer
annealing at 56 C for 20 s and extension at 72 C for 30 s; and a
final extension step at 72 C for 20 min.