Assessment of PowerPlex® Fusion 5C’s ability to type degraded DNA |
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| Institution: | 1. Henry C. Lee College, University of New Haven, Dodds Hall, 300 Boston Post Road, West Haven, CT 06516, USA;2. Connecticut-DESPP, Division of Scientific Services, 278 Colony Street, Meriden, CT 06451, USA;3. College of Agricultural Sciences & Natural Resources, University of Nebraska-Lincoln, 103 Agriculture Hall, Lincoln, NE 68583-0702, USA;1. Friendly Foot Care, PC, 50 West 94th Place, Crown Point, IN 46307, USA;2. St. Vincent Hospital, 123 Summer Street, Worcester, MA 01608, USA;3. University of Northampton, University Drive, Northampton NN1 5PH, UK;4. University of Northampton, University Drive, Northampton NN1 5PH, UK;1. University of Brescia - Department of Medical and Surgical Specialties, Radiological Sciences and Public Health, Forensic Medicine Unit, Brescia, Italy;2. Politec/MT - Forensic DNA Laboratory - Cuiaba, Mato Grosso, Brazil;1. Institute of Legal Medicine, Medical University of Innsbruck, Innsbruck, Austria;2. Forensic Science Program, The Pennsylvania State University, PA, USA;1. Criminal Science & Technology Institute, Bureau of Public Security, Urumqi, China;2. Ministry of Education Key Laboratory of Contemporary Anthropology and State Key Laboratory of Genetic Engineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Fudan University, Shanghai, China |
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| Abstract: | DNA samples collected at crime scenes are often degraded so this research focused on the ability of the Promega PowerPlex® Fusion 5C amplification kit to type both naturally and artificially degraded DNA.DNA was degraded naturally by placing equal volumes of blood on white fabric that was stored either inside, outside in a shaded area, or outside in direct sunlight. Samples were then collected every 10 days for 60 days and the DNA extracted (QIAamp® DNA Investigator). Artificially degraded samples were created by exposing extracted DNA to either UV light or 95 °C heat for varying times. DNA was also degraded artificially by placing blood samples into a 50% bleach solution for varying times prior to extraction.Following sample treatment, standard forensic DNA analysis was performed including quantification (Investigator® Quantiplex) and amplification (PowerPlex® Fusion 5C). Separation and detection were performed on an ABI 3130xl capillary electrophoresis unit and analysis was performed using GeneMapper ID v3.2.1.While the time and shade samples showed similar amounts of degradation, the samples exposed to direct sun showed more degradation. The artificially degraded samples showed more signs of degradation such as reduced overall peak height and peak height imbalance at heterozygous loci. There were also some cases where an allele that was known to be in the profile exhibited total dropout. Although there were some instances of both allelic dropout and heterozygote peak imbalance, PowerPlex® Fusion was able to reliably type degraded DNA as all alleles detected were consistent with the known donor profile. The results show that PowerPlex® Fusion is a robust kit capable of handling forensically challenged samples. |
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| Keywords: | Forensic Science Degraded DNA PowerPlex® Fusion 5C UV light Environmental Degradation Bleach treatment |
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