The 3-day international training course “Ancient DNA extraction course” organised for the forensic researchers from Italy was performed in the Laboratory of Molecular Genetics at the Institute of Forensic Medicine, Faculty of Medicine, University in Ljubljana in October 2017, under organization and leadership of project leader. The training course included experimental individual work with ancient bones and provided the participants first-hand knowledge of how to perform bone DNA typing. Procedures for processing the bone sample (mechanical and chemical cleaning, cutting, and grinding into the powder in presence of liquid nitrogen), decalcification of bone powder, DNA extraction, DNA purification, DNA quantification with real-time PCR, DNA typing of nuclear STRs, electrophoretic separation of amplified fragments and evaluation of DNA typing results were shown on concrete ancient bone samples and the most of the steps were experimentally performed by participants. The course was designed to deliver advanced level training to experienced laboratory based scientists that are familiar to DNA typing technologies. The unique training course was performed in the forensic molecular genetic laboratory equipped specially for processing old bones and teeth.
F.18 Transfer of new know-how to direct users (seminars, fora, conferences)
COBISS.SI-ID: 33490649The Prešeren's research, which was awarded by the Faculty of Medicine with the Prešeren’s prize for 2017 was performed in the Laboratory of Molecular Genetics at the Institute of Forensic Medicine, Faculty of Medicine, University in Ljubljana, under mentorship of project leader. The aim of this study was to determine the suitability of small bones of the hands and feet (metacarpals, metatarsals and phalanges) for genetic typing alongside the currently recommended femur, teeth, and the temporal bone. Three hypotheses were set (1. the amount of preserved ancient DNA extracted from the temporal bones, teeth and femurs can be compared to the amount extracted from small bones of the hands and feet (metacarpal bones, metatarsal bones, phalanges), 2. the success rate of autosomal STR typing from ancient DNA extracted from the temporal bones, teeth and femurs can be compared to the rate from small bones of the hands and feet, and 3. the success rate of autosomal STR typing has a strong positive relationship with the amount of extracted ancient DNA). We included 93 bone samples from 13 different skeletons obtained from archaeological sites in Slovenia. Six different skeletal elements were selected from each of the skeletons (temporal bone, teeth, femur, metacarpal bones, metatarsal bones, phalanges). We successfully obtained full or partial genetic profiles from at least one sample per skeleton while half of the samples weren’t eligible for genetic typing. The results confirmed all 3 of our hypothesis. The amount of preserved ancient DNA extracted from the temporal bones, teeth and femurs was comparable to the amount extracted from small bones of the hands and feet. The success rate of autosomal STR typing was comparable between the selected skeletal elements, and had a strong positive relationship with the amount of extracted ancient DNA. These findings suggest it would be appropriate to expand the sampling of skeletal elements for genetic typing of old skeletal remains and include the small bones of the hands and feet. To substantiate the findings of this study further research is needed using a larger amount of skeletons and small bones of the hands and feet.
E.01 National awards
COBISS.SI-ID: 3898388For extracting genomic DNA from Second World War (WWII) victim’s remains, a highly efficient extraction protocol was developed in our laboratory and presented on the conference in Portugal. We analysed more than 150 bones and teeth from WWII mass graves to evaluate this method. Procedures for processing the bone and tooth samples including mechanical and chemical cleaning, cutting and powdering in the presence of liquid nitrogen, complete demineralization of bone and tooth powder, DNA extraction, DNA purification using magnetic beads, DNA quantification and typing, were shown. The most appropriate types of bones and teeth for genetic analyses were described and the measures for preventing contamination in the DNA laboratory were listed. Some examples of identification of the Second World War mass grave victims were presented, focused mainly on DNA identification of skeletal remains of 88 victims of World War II killings found in the Konfin I mass grave. Living relatives were traced for 44 victims of Konfin I mass grave and genetic profiles of 84 right femurs were compared to the genetic material of living relatives. Since close and distant relatives (brothers, sisters, sons, daughters, nephews, or cousins) were available for identification of Konfin I victims, autosomal, paternaly inherited Y-chromosomal and maternaly inherited mitochondrial DNA (mtDNA) typing were performed and complete genetic profiles of autosomal, Y-chromosomal and mtDNA were obtained from almost all bone samples, allowing genetic identification of 32 victims of Konfin I. mass grave.
B.03 Paper at an international scientific conference
COBISS.SI-ID: 33673177