Dealing with low amounts of degraded DNA: Evaluation of SNP typing of challenging forensic samples by using massive parallel sequencing |
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| Institution: | 1. Department of Biomedical Sciences and Public Health, Section of Legal Medicine, Polytechnic University of Marche, Ancona, Italy;2. Department of Medical and Surgical Sciences, University of Bologna, Italy;3. Department of Public Health, Experimental and Forensic Medicine, University of Pavia, Italy;4. Section of Legal Medicine and Forensic Science, S. Maria Hospital, University of Perugia, Terni, Italy;5. Department of Public Health Sciences and Pediatrics, University of Turin, Turin, Italy;6. Department of Medicine, Surgery and Health, University of Trieste, Italy;7. Department of Drug Sciences, University of Pavia, Italy |
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| Abstract: | A set of eighty-two forensic samples with different levels of degradation, as well five in vitro damaged samples were analyzed by the Precision ID Identity Panel. PCR amplifications were performed with scalar amount of DNA (from 1 ng to 12 pg) and through different number of cycles. A minimum coverage of 50 x was adopted for “locus call”. Very informative profiles (based on about 65–70% of the loci) were obtained even in highly degraded samples when the amount of template range from 0.1 to 1.0 ng. When dealing with low amount of degraded DNAs, no more than half of the loci were typed, and the risk of mistyping (due to drop out phenomena) increased dramatically. The employment of a high number of PCR cycles is discussed. |
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| Keywords: | Degraded DNA MPS SNPs Allelic drop-out |
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