Global FilerTM PCR Amplification Kit is a highly discriminative STR kit developed by Life Technologies to meet the requirements of FBI recommendations, including 21 autosomal STR loci (i.e., D3S1358, vWA, D16S539, CSF1PO, TPOX, D8S1179, D21S11, D18S51, D2S441, D19S433, TH01, FGA, D22S1045, D5S818, D13S317, D7S820, SE33, D10S1248, D1S1656, D12S391 and D2S1338), 1 Y STR locus (i.e., DYS391), 1 Y-indel locus and Amelogenin. In this study, we did population genetic analyses with the GlobalFilerTM Kit for Uzbek population. 1380 samples were collected, with consent, from unrelated individuals from 12 geographic regions of Uzbekistan and Nukus region of Karakalpakistan. The samples were anonymized prior to analysis. Genomic DNA was extracted from buccal swabs using Swab SolutionTM Kit (Promega) protocol and amplified using the Global Filer TM PCR Amplification Kit (Applied Biosystems, USA) in the Veriti 96 Well Thermal Cycler (Applied Biosystems, USA), Proflex 3×32 well PCR System (Thermo Fisher Scientific, Singapore)and Proflex 96 well PCR System (Thermo Fisher Scientific, Singapore)according to the manufacturer’s recommendations. The electrophoretic separation of the amplified PCR products was performed on the ABI 3130xl Genetic Analyser (Applied Biosystems, USA). GeneScan-600 LIZ was used as the internal lane standard. The data analysis and allele identification were performed using GeneMapper ID–X (Applied Biosystems, USA) (version 1.4) analysis software (Life Technologies).
Statistical analysis such as observed heterozygosity (Ho), expected heterozygosity (He), Match Probability (MP), heterozygosity (H), the power of discrimination (PD), power of exclusion (PE), Paternity Index (PI), and polymorphism information content (PIC) were calculated using Power Stats v 1.2 Promega Corporation. The allele frequencies of samples and exact tests of Hardy-Weinberg equilibrium (HWE) were calculated using Genepop v 4.7.0.
All loci are polymorphic in all populations. Y-indel is the least informative marker as it has only two alleles. After Bonferroni correction (p-value=0.05/21=0.0024 for HWE test and 0.05/21=0.00024 for LD test), no locus was departed from HWE and no pair of loci were departed from LD.
An MDS plot, based on Fst distances, of the Uzbek populations and four US major populations (i.e., Caucasian, African American, Hispanic, and Asian) are generally consistent with geographical distances or population admixture history.
This study supports that the autosomal STR loci amplified with the Global FilerTM Kit have high discrimination powers in Uzbek population. The allele frequencies can be used in forensic and paternity testing cases.