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Online ISSN 1827-1677
Fabbri M., Venturi M., Ferronato C., Tarantino S., Gaudio R. M., Avato F. M.
Laboratory of Forensic Genetics Section of Legal Medicine Department of Medical Sciences University of Ferrara, Ferrara, Italy
Aim: Our work involved the extraction, PCR amplification, and the definition of the genetic profile of 5 skeletal elements, dating back to World War II and delivered to the laboratory by two associations of victims of World War II operating in the province of Ferrara.
Methods: First, bone fragments have been treated in order to remove residual soil. Subsequently, all the bone fragments were treated with UV light to remove exogenous DNA may be present on the outer surface. After this preliminary phase, it was collected about 5g of tissue, represented by both compact and soft tissue, after removal of the outermost layers. The samples collected were treated, prior to DNA extraction, using 25-30ml of 0.5 M EDTA, pH 8.0-8.5, monitoring the process by the addition of ammonium oxalate at pH 4.0. After washing with 25ml of sterile distilled water and centrifugation at 4000rpm for 20 minutes, DNA was extracted from pellets. After digestion process, the tubes containing the extracts were placed at -20 °C for 24 hours. The method, therefore, predicted n. 2 cycles of purification in phenol-chloroform-isoamyl alcohol (25:24:1) Because of bone samples were particularly dated, DNA was concentrated using Centricon YM-30 . DNA concentration was determined using a spectrophotometer BioPhotometer (Eppendorf).
Results: The analysis conducted at our laboratory allowed to verify the absence of contamination by exogenous DNA, both in human remains used in the analysis both inside the solutions used for extraction and amplification. The analysis of the electrophoretic patterns showed n. 5 partial genetic profiles (6-9 STR markers), referring to n. 5 unrelated male.
Conclusion: These first our results could be used as an internal laboratory model for further DNA identification of human remains referred to World War II mass that are, unfortunately, scattered in our country region.