ABSTRACT
IndroductionThe short tandem repeats (STR) are rich source of highly polymorphic markers in the human genome. In this study, we used a commercially available multiplex STR typing kit to study 15 STR systems (D3S1358, THO1, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSF1PO, Penta D, vWA, D8S1179, TPOX and FGA,) in the Mongolians population, and estimated the allele and genotype frequencies. These 15 STR loci include 2 new pentanucleotide repeat STR loci, Penta E and Penta D, so are not studied in Mongolians.GoalTo determine allele frequency of STR loci D3S1358, THO1,D21S11, D18S51, D5S818, D13S317, D7S820, D16S539, CSF1PO, vWA, D8S1179, TPOX, FGA Penta E, Penta D in Mongolian population.Materials and MethodsThe liquid blood, blood stain and saliva samples were taken from 165 unrelated individuals from Mongolian. Extraction DNA: Genomic DNA was extracted from whole blood samples by the standard method of phenol-chloroform-isoamyl alcohol and Wizard Genomic DNA Purification kit, Promega Corporation [21], from blood stain and saliva samples QIAamp DNA micro kit, Qiagen [25], AccuPrep Genomic DNA Extration kit, Bioneer, Koreans extraction method respectively.PCR: PCR amplification was performed using 10-15 ng genomic DNA template according to manufacturer’s protocol (PowerPlex® 16 and PowerPlex® 16HS kit, Promega Corporation, USA). Typing: DNA typing was performed on the ABI Prism 310 Genetic Analyzer (Applied Biosystems) using the recommended protocol. The results were analyzed by Data Collection (Version 1.1), GeneScan (Version 3.1), and Genotyper (Version 3.1) softwares (AppliedBiosystems).ResultsWe assessed forensic and population genetic studies using 15 STR loci included in s sample of 165 unrelated individuals from Mongolian. Allele frequency were listed in Table 2. Totally 20 alleles /5, 7-25/ were found from microsatellite Penta E locus and allele 11 has most frequent (0.1128). 6-16 alleles were found from Penta D locus and allele 9 has most frequent (0.3262). This result is interesting because allele 6 of Penta D locus was found rarely among other populations. But relatively higher frequency of allele 6 (0.0183) was found in Mongolian population. A population comparison based in genetic distance and genetic diversity calculated from allele frequencies of the 15 STR loci from obtained five different populations is shown the Table 3. Conclusions:1. Penta E locus was highly polymorphic, and 20 alleles were found in this Mongolians population and allele 11 was most frequent.2. Penta D locus was 20 alleles were found in this Mongolians population and allele 9 was most frequent.
ABSTRACT
DNA quantification is important to ensure the consistency and the reliability in the interpretation of degraded low copy number DNA typing. We applied the simple PCR quantification method using fluo-rescently labeled primers for the amplification of mtDNA and amelogenin gene in 50 year old skeletal remains (e.g. bone and tooth). K562 DNA was serially diluted and used as a standard for concentration marker to gauge the amount of DNA from PCR versus the peak area. The quantities of DNA extracted from bones and teeth did not show significant difference in the analyses both using mtDNA and amelo-genin gene as an amplification target. To test the efficiency of DNA profiling of degraded low copy number DNA samples, mtDNA PCR quality evaluation and DNA typing for 16 autosomal STR and 9 Y chromosomal STR loci were per-formed and the correlation between DNA quantities and PCR amplification efficiencies of the samples was analyzed. The DNA quantities assayed by the simple method suggested in the present study could be good indicator for mtDNA and STR analysis. As the allele drop-out was observed in less than 0.050ng DNA samples, at least 0.100ng of DNA is required to produce informative STR profiles. Also, STRs with less than 200bp amplification sizes produce efficient DNA profiles in most cases. Therefore, the develop-ment of mini-STRs with less than 200bp amplification sizes is expected to improve DNA typing in degraded low copy number DNA. Y-STRs are easy to detect allele drop-out or drop-in, and accordingly the efficiency test of Y-STRs as well as autosomal STRs for profiling of degraded low copy number DNA samples is thought to be important.