Whole blood obtained by venipuncture was collected in EDTA vacutainer tubes from individuals attending Split Hospital, Split,Croatia. Blood was dried onto clean cotton cloth, sealed in individual plastic bags and transported to the United States for testing. Approximately 3 mm squares were extracted using Chelex (1). The DNA was not quantified. PCR amplification was performed using the AmpFlSTR Profiler Plus™ and Cofiler™ PCR amplification kits (PE-Biosystems, Foster City, CA) following the manufacturer's protocol using 2 µL of sample. The amplified products were separated and detected using the ABI Prism™ 377 DNA sequencer (PE-Biosystems, Foster City, CA). Allele frequencies and goodness of fit tests were performed using Excel spread sheets. Each locus (n = 13) was tested for the following: Hardy-Weinberg Global Chi-square goodness of fit using all genotypes with expected values greater than 1.0, all other genotypes were pooled to form a residual class (df = number of genotypes minus the number of alleles)( p1); T statistic testing θ = 0 recommended by NRC 2 (2)(p2); and the Chi-square comparing expected and observed total homozygosity and heterozygosity (df = 1)(p3). The Bonferroni corrected threshold was 0.0038. No loci exceeded the Bonferroni threshold; only one locus (D8S1179) was significant (p (0.05) for one of three tests of deviation from Hardy-Weinberg equilibrium due to an excess of heterozygotes and deficiency of homozygotes prior to the Bonferroni correction for multiple tests.
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