Allele Frequencies for 15 Short Tandem Repeat Loci in Representative Sample of Croatian Population

Aim To study the distribution of allele frequencies of 15 short tandem repeat (STR) loci in a representative sample of Croatian population. Methods A total of 195 unrelated Caucasian individuals born in Croatia, from 14 counties and the City of Zagreb, were sampled for the analysis. All the tested individuals were voluntary donors. Buccal swab was used as the DNA source. AmpFlSTR® Identifiler® was applied to simultaneously amplify 15 STR loci. Total reaction volume was 12.5 μL. The PCR amplification was carried out in PE Gene Amp PCR System Thermal Cycler. Electrophoresis of the amplification products was preformed on an ABI PRISM 3130 Genetic Analyzer. After PCR amplification and separation by electrophoresis, raw data were compiled, analyzed, and numerical allele designations of the profiles were obtained. Deviation from Hardy-Weinberg equilibrium, observed and expected heterozygosity, power of discrimination, and power of exclusion were calculated. Bonferroni’s correction was used before each comparative analysis. Results We compared Croatian data with those obtained from geographically neighboring European populations. The significant difference (at P<0.01) in allele frequencies was recorded only between Croatian and Slovenian populations for vWA locus. There was no significant deviation from Hardy-Weinberg equilibrium for all the observed loci. Conclusion Obtained population data concurred with the expected “STR data frame” for this part of Europe

Optimisation of Forensic Genetics Procedures Used in Disputed Paternity Testing: Adjustment of the PCR Reaction Volume

Standard molecular techniques, with only a slight modification, are very useful in obtaining and interpreting the final results in the field of forensic genetic. Data obtained through such analysis are highly reliable and can be used as a very powerful tool that produces valuable results. However, success and swiftness of DNA typing of biological evidence either that found at a crime scene or used in disputed paternity testing, depends on the optimization of numerous factors. One of the most important and critical phases that ensures reliability of the whole procedure is the choice of the most suitable volume for the amplification protocol. Buccal swabs were collected from volunteers. DNA was extracted by Qiagen Dnaeasy Tissue Kit. PowerPlex 16 kit was used to simultaneously amplify 15 STR loci by PCR. Amplification was carried out as described previously. The tested total working reaction volumes were 5, 10 and 25 microl. The PCR amplification was carried out in PE Gene Amp PCR System Thermal Cycler (ABI, Foster City, CA). Amplification products were analyzed on an ABI PRISM 377 instrument (ABI, Foster City, CA) in 5% bis-acrilamide gel. Amplification was generally successful for all the tested reaction volumes. Lower partial to complete DNA profiles ratio, the quality of obtained STR profiles, significantly reduced amount of reaction's components give advantage to 5 microl reaction volume over other two tested volumes in this case

Allele Frequencies for 15 Short Tandem Repeat Loci in Representative Sample of Croatian Population

Aim To study the distribution of allele frequencies of 15 short tandem repeat (STR) loci in a representative sample of Croatian population. Methods A total of 195 unrelated Caucasian individuals born in Croatia, from 14 counties and the City of Zagreb, were sampled for the analysis. All the tested individuals were voluntary donors. Buccal swab was used as the DNA source. AmpFlSTR® Identifiler® was applied to simultaneously amplify 15 STR loci. Total reaction volume was 12.5 μL. The PCR amplification was carried out in PE Gene Amp PCR System Thermal Cycler. Electrophoresis of the amplification products was preformed on an ABI PRISM 3130 Genetic Analyzer. After PCR amplification and separation by electrophoresis, raw data were compiled, analyzed, and numerical allele designations of the profiles were obtained. Deviation from Hardy-Weinberg equilibrium, observed and expected heterozygosity, power of discrimination, and power of exclusion were calculated. Bonferroni’s correction was used before each comparative analysis. Results We compared Croatian data with those obtained from geographically neighboring European populations. The significant difference (at P<0.01) in allele frequencies was recorded only between Croatian and Slovenian populations for vWA locus. There was no significant deviation from Hardy-Weinberg equilibrium for all the observed loci. Conclusion Obtained population data concurred with the expected “STR data frame” for this part of Europe

Most recent investigation of peopling of Bosnia and Herzegovina: DNA approach

Many historical episodes marked Bosnia and Herzegovina as a significant ethnic crossroads, which makes it a very interesting site for various population studies. The first stages of these complex investigations were based on observations of numerous phenotype markers. The following phase, which was relatively brief, was dominated by the use of different cytogenetic markers. Finally, at the beginning of this century, the molecular-genetic diversity of the BiH population became the focus of modern research. Autosomal and Y-STR markers, together with mitochondrial haplogroup (Hg) diversity were initially used in the examination of isolated groups, as well as the whole population of modern Bosnia and Herzegovina. The most recent study describes the distribution of Y-chromosome haplogroups in the three main ethnic groups in Bosnia and Herzegovina, and suggests a preliminary hypothesis for the process of peopling this area.Veliko zgodovinskih dogodkov je zaznamovalo Bosno in Hercegovino kot pomembno etnično stičišče, ki je zelo zanimivo za različne populacijske študije. Začetki kompleksnih raziskav so povezani z opazovanjem številnih fenotipskih označevalcev. V naslednji fazi je prevladala uporaba različnih citogenetskih označevalcev. Na začetku tega stoletja so raziskave usmerjene v molekularno-genetsko raznolikost BiH populacije. Pri raziskavah izoliranih populacij, kot tudi celotne človeške populacije moderne Bosne in Hercegovine, so na začetku uporabili avtosomalne in Y-STR označevalce skupaj z raznolikostjo mitohondrijske haploskupine (Hg). Najnovejša študija opisuje porazdelitev haploskupin Y-kromosoma pri treh glavnih etničnih skupinah v Bosni in Hercegovini in izdela preliminarni scenarij procesa poselitve tega področja