Development of a quantitative PCR-based method for studying temporal DNA degradation in waterlogged bone
| Autor: | Claire Cartozzo, Darcy P. Mays, Baneshwar Singh, Isis Thornton, Tal Simmons |
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| Rok vydání: | 2020 |
| Předmět: |
Veterinary medicine
Swine Fresh Water Ribs Biology Real-Time Polymerase Chain Reaction Pathology and Forensic Medicine Submersion (mathematics) chemistry.chemical_compound DNA degradation Immersion Genetics TaqMan Animals DNA Degradation Necrotic DNA DNA extraction DNA Fingerprinting Scapula genomic DNA Real-time polymerase chain reaction chemistry Aquatic environment Models Animal Microsatellite Repeats |
| Zdroj: | Journal of forensic sciencesREFERENCES. 66(3) |
| ISSN: | 1556-4029 |
| Popis: | While several studies have examined temporal DNA degradation in bones collected from terrestrial environments, studies on temporal DNA degradation in bones collected from aquatic environments are limited and mostly based on case studies. The objective of this study was to assess the impact of long-term submersion, aquatic environment, bone type and DNA extraction method on DNA quality and quantity. Bone samples (scapulae and ribs), collected every ~1000 ADD from a freshwater lake and river, underwent DNA extraction via ChargeSwitch® gDNA Plant Kit and organic phenol-chloroform methods, and DNA quantitation using both TaqMan and SYBR Green-based quantitative PCR (qPCR) methods. Results suggest that in both bone types, quality of recovered DNA (i.e., degradation index) declined significantly with increase in submersion time. Among two bone types, quality of recovered DNA from scapulae declined faster than rib samples. There was no significant difference in recovered DNA quantity between bone types, DNA extraction methods, or locations but various interactions between these variables showed significant difference. Overall, it can be concluded that DNA can be extracted from waterlogged bone in sufficient quantity to generate an STR profile up to 4000 ADD. |
| Databáze: | OpenAIRE |
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