Published Online: 1 November 2003
Page Count: 2
Genomic DNA was obtained from peripheral blood extracted with Chelex-100 method (1). PCR was performed in a total reaction volume of 20 µL containing 20 ng genomic DNA, 0.2 µM each primer, 10 mM Tris-HCl buffer (pH 8.3), 50 mM KCl, 1.5 mM MgCl2, 200 RM each dNTP, and 1 U Taq DNA polymerase (BioStar, Canada). Primer sequences: D2S1788: 5 -aat gga tgg aca aat gga tg-3, 5 -ccc tcc ata att aga tga gcc-3; D6S1043: 5 -caa gga tgg gtg gat caa ta-3, 5 -ttg tat gag cca ctt ccc at-3; D7S3048: 5 -ctg gag ctg cat agt gtc ct-3, 5 -aat cat ccc tgt gtg ctt tc-3; D12S1064: 5 -act act cca agg ttc cag cc-3, 5 -aat att gac ttt ctc ttg cta ccc-3. PCR conditions: 95°C for 2 min soak, 32 cycles of 35 s at 94°C, 35 s at 60°C, 40 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. Allele frequencies and others statistics parameters for forensic and paternity were determined for each locus by the PowerStats software packages (2). The Hardy-Weinberg equilibrium test (HWE) was performed by an exact test (3). None of the analyzed loci showed deviations from HWE (P > 0.05) in the population studied. The complete dataset is available to any interested researcher upon request to the authors.
Paper ID: JFS2003201