“…These microsatellite loci were amplified by polymerase chain reaction (PCR) using a Veriti thermal cycler (Applied Biosystems, Foster City, CA, USA) with a total volume of 10 µL per reaction (containing 10 ng of genomic DNA, 1× buffer, 210 µM of each dNTP, 1.5 mM MgCl 2 , 0.16 µM forward primer and M13 (FAM or NED) broth [23], 0.32 µM reverse primer, 1.05 U Taq DNA polymerase (Invitrogen, Carlsbad, CA, USA), and 3.49 µL ultrapure water. These PCR amplifications consisted of two steps, of which the first was primer-specific and the second M13 binding [24]. The amplification products were checked by electrophoresis on 1.5% agarose gels stained with GelRed (Biotium, Fremont, CA, USA) in 1× TBE buffer (pH 8.0).…”