Low-cost direct PCR for aged and processed wildlife sample analysis |
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| Institution: | 1. Forensic Science Program, Department of Applied Science, Faculty of Science, Prince of Songkla University, Thailand;2. School of Biological Sciences, Flinders University, Adelaide, South Australia, Australia;3. Princess Maha Chakri Sirindhorn Natural History Museum, Prince of Songkla University, Thailand;4. Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla University, Thailand;1. Institute of Biology and Medical Genetics of the First Faculty of Medicine and General University Hospital in Prague, Charles University in Prague, Czech Republic;2. Institute of Forensic Medicine and Toxicology of the First Faculty of Medicine and General University Hospital in Prague, Charles University in Prague, Czech Republic;3. Institute of Criminalistics Prague, Police of the Czech Republic, Czech Republic;1. Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark;2. IPATIMUP–Institute of Molecular Pathology and Immunology of the University of Porto, Portugal;3. Faculty of Sciences of the University of Porto, Portugal;4. DNA Diagnostic Laboratory, State University of Rio de Janeiro, Brazil;1. Dipartimento di Scienze Biomediche e Sanità Pubblica, Sezione di Medicina Legale, Università Politecnica delle Marche, Ancona, Italy;2. Dipartimento di Sanità Pubblica, Neuroscienze, Medicina Sperimentale e Forense, Sezione di Medicina Legale e Scienze Forensi, Università di Pavia, Italy;3. Institute of Legal Medicine, Faculty of Medicine, University of Cologne, Germany;4. Dipartimento di Scienze della Salute, Sezione di Medicina Legale, Università di Genova, Italy |
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| Abstract: | Direct PCR has been used successfully in wildlife forensic DNA analysis from several types of biological samples using specialized, commercial direct PCR kits. This is attributed to the proprietary chemicals provided in the kits such as pre-PCR buffer and modified DNA polymerases. These reagents can be expensive, thereby limiting their widespread adoption in developing countries, where wildlife crimes are often encountered. We report on a study to evaluate the possibility of using low-cost direct PCR assay for degraded and processed wildlife sample analysis. Phire® and Q5® polymerases were used, due to their relatively low cost, for direct amplification of six aged and processed sample types (dried skin, 30-year old hair, muscle tissue, bone, trace blood mixed in vodka, and dried soft antler). The result indicated that Phire® Hot Start II DNA polymerase and Q5® DNA polymerase performed similarly to commercially available direct PCR kit. The low-cost amplification could efficiently identify species origin from all aged and processed samples. We observed a rate of more than 80% amplification success and high PCR product concentrations sufficient for further sequencing. The assay proved to be cost effective and robust; thus, we expect it to be adopted by wildlife forensic laboratories in developing countries. |
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| Keywords: | Direct PCR Low-cost Aged Processed Wildlife forensic science |
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