Abstract
EDTA whole blood samples were collected from 281 unrelated individuals of Han population living in Wuhan, China. DNA was extracted using Cheles-100 method (1). Hot-start PCR was performed in a total volume of 10 µL containing 10 ng genomic DNA, 0.2 µM each primer, 10 mM Tris-HCl buffer (pH8.3), 50 mM KCl, 1.5 mM MgCl2, 200 µM each dNTP, and 0.3 U Taq DNA polymerase (BioStar, Canada) was add when the temperature reaches 92°C. The primer sequences newly designed by us were: Penta D: 5'-cagagcaagacaccatctcaa-3', 5'-tttgcctaacctatggtcataacg-3'; Penta E: 5'-agatcacgccattgcactcc-3', 5'-gggttattaattgagaaaactccttacaat-3'. PCR cycling conditions: 95°C for 2 min soak, 32 cycles of 30 s at 94°C, 30 s at 62°C for Penta D and 60°C for Penta E, 35 s at 72°C followed by a 5 min extension period at 72°C. The amplification products were separated in a vertical, native polyacrylamide gel (6% T; 5% C) and visualized by silver staining (2). Allele frequencies and other statistics parameters for forensic and paternity were determined for these two locus by the PowerStats software packages (3). The Hardy-Weinberg equilibrium test (HWE) was performed by an exact test (4). None of the analyzed loci showed deviations from HWE (P > 0.05) in the population studied.
