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.

The variation at 28 Y‐chromosome biallelic markers was analysed in 256 males (90 Croats, 81 Serbs and 85 Bosniacs) from Bosnia‐Herzegovina. An important shared feature between the three ethnic groups is the high frequency of the “Palaeolithic” European‐specific haplogroup (Hg) I, a likely signature of a Balkan population re‐expansion after the Last Glacial Maximum. This haplogroup is almost completely represented by the sub‐haplogroup I‐P37 whose frequency is, however, higher in the Croats (∼71%) than in Bosniacs (∼44%) and Serbs (∼31%). Other rather frequent haplogroups are E (∼15%) and J (∼7%), which are considered to have arrived from the Middle East in Neolithic and post‐Neolithic times, and R‐M17 (∼14%), which probably marked several arrivals, at different times, from eastern Eurasia. Hg E, almost exclusively represented by its subclade E‐M78, is more common in the Serbs (∼20%) than in Bosniacs (∼13%) and Croats (∼9%), and Hg J, observed in only one Croat, encompasses ∼9% of the Serbs and ∼12% of the Bosniacs, where it shows its highest diversification. By contrast, Hg R‐M17 displays similar frequencies in all three groups. On the whole, the three main groups of Bosnia‐Herzegovina, in spite of some quantitative differences, share a large fraction of the same ancient gene pool distinctive for the Balkan area.

All 193 tested individuals have been involved in legal proceedings concerning various forensic testing. Buccal swabs have been taken as the DNA source and Chelex procedure was used for DNA extraction (1). The PowerPlex 16 kit (Promega Corp., Madison, WI) has been used to simultaneously amplify by PCR 15 STR loci. The STR loci are: D3S1358, TH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSF1PO, Penta D, vWA, D8S1179, TPOX and FGA. Similar amounts of DNA have been used in all PCR reactions. Amplification was carried out as described previously (2).

POPULATION: We have analyzed the distribution of allele frequencies at 12 Y-chromosornal short tandem repeats loci (DYS 19, DYS385a, DYS385b. DYS389I. DYS389II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438 and DYS439) in the representative sample of Bosnian and Herzegovinians. A total of 100 unrelated male individuals (Caucasians) from different regions of Bosnia and Herzegovina have been sampled for the analysis. Samples were collected with a respect to the approximate proportional participation of three main ethnic groups in Bosnia and Herzegovina [Bosniacs-Muslim (35). Serbs (31), Croats (34)].

The debate concerning the mechanisms underlying the prehistoric spread of farming to Southeast Europe is framed around the opposing roles of population movement and cultural diffusion. To investigate the possible involvement of local people during the transition of agriculture in the Balkans, we analysed patterns of Y-chromosome diversity in 1206 subjects from 17 population samples, mainly from Southeast Europe. Evidence from three Y-chromosome lineages, I-M423, E-V13 and J-M241, make it possible to distinguish between Holocene Mesolithic forager and subsequent Neolithic range expansions from the eastern Sahara and the Near East, respectively. In particular, whereas the Balkan microsatellite variation associated to J-M241 correlates with the Neolithic period, those related to E-V13 and I-M423 Balkan Y chromosomes are consistent with a late Mesolithic time frame. In addition, the low frequency and variance associated to I-M423 and E-V13 in Anatolia and the Middle East, support an European Mesolithic origin of these two clades. Thus, these Balkan Mesolithic foragers with their own autochthonous genetic signatures, were destined to become the earliest to adopt farming, when it was subsequently introduced by a cadre of migrating farmers from the Near East. These initial local converted farmers became the principal agents spreading this economy using maritime leapfrog colonization strategies in the Adriatic and transmitting the Neolithic cultural package to other adjacent Mesolithic populations. The ensuing range expansions of E-V13 and I-M423 parallel in space and time the diffusion of Neolithic Impressed Ware, thereby supporting a case of cultural diffusion using genetic evidence.