Amplification was carried out with
a 10 × PCR buffer (100 mM Tris-HCl, 1 mM DTT, 0.1 mM
EDTA, 100 mM KCl, 0.5% Nonidet P40, 0.5% Tween 20)
in a total reaction of 50 μl containing 2.5 mM dNTP, 20 mM
MgCl2, 100 pmol of primer, 2 μl of genomic template DNA,
and 1 unit of Taq DNA polymerase (DNA Gdansk, Poland).