Daniel Vanek

Czech population data on 10 short tandem repeat loci of SGM Plus STR system kit using DNA purified in FTA™ cards

A population study on 10 short tandem repeat (STR) loci was performed on 202 unrelated Czech Caucasians. DNA, initially frozen, was thawed and placed FTA paper. The DNA was purified in situ. DNA was amplified by PCR using the AmpFlSTR SGM Plus amplification kit.

Technical note: Efficient removal of a PCR inhibitory agent (vivianite) found on excavated bones.

During a rescue excavation in October 2011, archaeologists discovered a mass grave with 10 individuals. The skeletons should belong to victims of the battle of Reichenberg between the Austrian and Prussian armies on April 21, 1757. Several bones of the skeletons were covered with a blue colored encrustation. Initial DNA analysis failed due to strong inhibition. Chemical analysis of the bluish encrustation indicated the presence of the iron phosphate mineral vivianite (Fe3(PO4)2·(H2O)8). This technical note describes a novel procedure for the removal of this inhibitory substance.

Results of a collaborative study on DNA identification of aged bone samples

Aim A collaborative exercise with several institutes was organized by the Forensic DNA Service (FDNAS) and the Institute of the Legal Medicine, 2nd Faculty of Medicine, Charles University in Prague, Czech Republic, with the aim to test performance of different laboratories carrying out DNA analysis of relatively old bone samples. Methods Eighteen laboratories participating in the collaborative exercise were asked to perform DNA typing of two samples of bone powder. Two bone samples provided by the National Museum and the Institute of Archaelogy in Prague, Czech Republic, came from archeological excavations and were estimated to be approximately 150 and 400 years old. The methods of genetic characterization including autosomal, gonosomal, and mitochondrial markers was selected solely at the discretion of the participating laboratory. Results Although the participating laboratories used different extraction and amplification strategies, concordant results were obtained from the relatively intact 150 years old bone sample. Typing was more problematic with the analysis of the 400 years old bone sample due to poorer quality. Conclusion The laboratories performing identification DNA analysis of bone and teeth samples should regularly test their ability to correctly perform DNA-based identification on bone samples containing degraded DNA and potential inhibitors and demonstrate that risk of contamination is minimized.

Improved Y-STR typing for disaster victim identification, missing persons investigations, and historical human skeletal remains

Bones are a valuable source of DNA in forensic, anthropological, and archaeological investigations. There are a number of scenarios in which the only samples available for testing are highly degraded and/or skeletonized. Often it is necessary to perform more than one type of marker analysis on such samples in order to compile sufficient data for identification. Lineage markers, such as Y-STRs and mitochondrial DNA (mtDNA), represent important systems to complement autosomal DNA markers and anthropological metadata in making associations between unidentified remains and living relatives or for characterization of the remains for historical and archaeological studies. In this comparative study, Y-STR typing with both Yfiler™ and Yfiler™ Plus (Thermo Fisher Scientific, Waltham, MA, USA) was performed on a variety of human skeletal remains, including samples from the American Civil War (1861–1865), the late nineteenth century gold rush era in Deadwood, SD, USA (1874–1877), the Seven Years’ War (1756–1763), a seventeenth-century archaeological site in Raspenava, Bohemia (Czech Republic), and World War II (1939–1945). The skeletal remains used for this study were recovered from a wide range of environmental conditions and were extracted using several common methods. Regardless of the DNA extraction method used and the age/condition of the remains, 22 out of 24 bone samples yielded a greater number of alleles using the Yfiler™ Plus kit compared to the Yfiler™ kit using the same quantity of input DNA. There was no discernable correlation with the degradation index values for these samples. Overall, the efficacy of the Yfiler™ Plus assay was demonstrated on degraded DNA from skeletal remains. Yfiler™ Plus increases the discriminatory power over the previous generation multiplex due to the larger set of Y-STR markers available for analysis and buffer modifications with the newer version kit. Increased haplotype resolution is provided to infer or refute putative genetic relationships.

Forensic genetic analyses in isolated populations with examples of central European Valachs and Roma

Isolated populations present a constant threat to the correctness of forensic genetic casework. In this review article we present several examples of how analyzing samples from isolated populations can bias the results of the forensic statistics and analyses. We select our examples from isolated populations from central and southeastern Europe, namely the Valachs and the European Roma. We also provide the reader with general strategies and principles to improve the laboratory practice (best practice) and reporting of samples from supposedly isolated populations. These include reporting the precise population data used for computing the forensic statistics, using the appropriate θ correction factor for calculating allele frequencies, typing ancestry informative markers in samples of unknown or uncertain ethnicity and establishing ethnic-specific forensic databases.