Letter to the Editor Allele frequencies of the new European Standard Set (ESS) loci plus SE33 locus in a population from the Republic of Macedonia Dear Editor, The highly polymorphic short tandem repeat (STR) loci in the human genome are widely used for forensic and paternity testing as well as for population genetic studies. Introducing set of STR loci as observed markers made marvels progress in this field of forensic science [1,2]. Nowadays, in response to the ENFSI and EDNAP groups’ call for new STR multiplexes for Europe, Promega developed a suite of four new DNA profiling kits. One of them is PowerPlex ESI 17 that we have used in this study. In our previous population studies of Macedonian human population, we have used 15 STR loci included in the AmpFl- STR 1 Identifiler 1 [3] and 17 Y-chromosomal short tandem repeats loci incorporated in the AmpFistr 1 Yfiler TM PCR Amplification Kit (data submitted to the FSI Genetics). All obtained results were included in Macedonian referent database. In order of future development of this database we have decide to analyze 6 additional new STR loci (D22S1045, D1S1656, D10S1248, D2S441, D12S391, and SE33). Some of these loci were described even 15 years ago [4,5]. This study is based on an observation of representative sample of Macedonia human population. A total of 205 unrelated individuals of which 105 males and 100 females, from different regions of Macedonia, participated in this study. All tested individuals have been voluntary donors appropriately informed about the research. Qiagen Micro TM Tissue Kit was used for DNA extraction from whole blood, Quantifiler 1 assay for quantification and PowerPlex ESI 17 1 System for amplification and detection (D18S51, D21S11, TH01, D3S1358, Amelogenin, D16S539, D2S1338, D1S1656, D10S1248, FGA, D8S1179, vWA, D22S1045, D19S433, D12S391, D2S441 and SE33). The total volume of each reaction was 12.5 ml. The PCR amplifications have been carried out in ABI 9700 Thermal Cycler according to the manufacturer’s recommendations. Electrophoresis of the amplification products was performed on an ABI 310 PRISM. Numerical allele designations of the profiles were obtained by processing with Gene Mapper 1 IDv 3.2 Software. Deviation from Hardy–Weinberg equilibrium, observed and expected heterozygosity, power of discrimination and power of exclusion were calculated. Also, we have compared Macedonian data with data obtained from other neighboring populations with available data for observed loci. Deviation from Hardy–Weinberg equilibrium [6], observed and expected hetero- zygosity [7], exact test of population differentiation [8] and pairwise Fst [9] were calculated within Powermarker v3.25 [10], since power of discrimination and power of exclusion within Microsoft1 Excel workbook template—PowerStats [11]. Allele frequencies and all calculated forensic parameters are shown in Supplementary Table 1. All data are available on http:// www.ingeb.ba/rebida/ESI17/macedonia.html. No statistically significant deviation (p < 0.05) from Hardy- Weinberg equilibrium was found except for D1S1656 and SE33 loci (Table 1). Heterozygosity excess has been detected for SE33 locus. Allele frequencies of observed six loci within Macedonian population have been compared with data obtained from geographically neighboring Southeast European populations such as Serbia and Bosnia Herzegovina (our unpublished data). After Bonferroni’s correction, statistical significance was considered as Forensic Science International: Genetics 6 (2012) e90–e92 Table 1 The distribution of allele frequencies and calculated forensic parameters for the Macedonian population over 5 new ESS loci and SE33 locus (n = 205). Allele D1S1656 D2S441 D10S1248 D12S391 SE33 D22S1045 2n 410 410 410 410 410 410 1 – – – – – – 6.3 – – – – 0.0024 – 9 – 0.0024 – – – – 10 – 0.1268 – – – 0.0024 11 0.1146 0.3365 – – – 0.1366 11.3 – 0.0951 – – – – 12 0.1683 0.0415 0.0268 0.0049 0.0024 0.0122 12.1 – 0.0039 – – – – 12.3 – 0.0024 – – 0.0254 – 13 0.0683 0.0390 0.2341 – 0.0122 – 13.2 – – – – 0.0073 – 14 0.0780 0.3073 0.3293 0.0024 0.0098 0.0537 14.2 – – – – 0.0073 – 15 0.1244 0.0439 0.1780 0.0512 0.0463 0.3634 15.3 0.0463 – – – – – 16 0.1366 0.0024 0.1829 0.0317 0.0366 0.3220 16.1 – – – – 0.0024 – 16.3 0.0390 – – – – – 17 0.0341 – 0.0439 0.1000 0.0659 0.0951 Contents lists available at ScienceDirect Forensic Science International: Genetics jou r nal h o mep ag e: w ww .elsevier .co m /loc ate/fs ig 1872-4973/$ – see front matter ß 2011 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.fsigen.2011.07.004