EDTA-blood samples were collected from 176 healthy unrelated males of Han population living in Wuhan, China. DNA was extracted using Cheles-100 method (1). PCR was performed in a total volume of 10 µL containing 2–5 ng genomic DNA, 0.2 µM each primer, 10 mM Tris-HCl buffer (pH 8.3), 50 mM KCl, 1.5 mM MgCl2, 200 µM each dNTP, and 1 U AmpliTaq Gold® DNA polymerase (Applied Biosystems, Foster City, CA). DYS459 and DYS464 were co-amplified in a duplex reaction and DYS385 was amplified in a singleples reaction. Primer sequences: DYS385: 5'-FAM-agcatgggtgacagagcta-3', 5'-gccaattacatagtcctcctttc-3'; DYS459: 5'FAM-caggtgaactggggtaaataat-3', 5'-gttgagcaacagagcaagactta-3'; DYS464: 5'-FAM-ctttgggctatgcctcagttt-3', 5'-gccatacctgggtaacagagagac-3'. PCR cycling conditions: 95°C for 11 min soak, 30 cycles of 40 s at 94°C, 40 s at 60°C, 50 s at 72°C followed by a 6 min extension period at 72°C. All loci were amplified in a GeneAmp PCR System 9700 (PE Applied Biosystems).
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