The modified method of two-step differential extraction of sperm and vaginal epithelial cell DNA from vaginal fluid mixed with semen |
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| Institution: | 1. Office of the Chief Forensic Scientist, Victoria Police Forensic Services Centre, 31 Forensic Drive, Macleod 3085, VIC, Australia;2. Department of Genetics, La Trobe Institute for Molecular Sciences, La Trobe University, Melbourne 3086, VIC, Australia;3. Department of Biochemistry, La Trobe Institute for Molecular Sciences, La Trobe University, Melbourne 3086, VIC, Australia;1. Department of Forensic Chemistry, College of Forensic Sciences, Naif Arab University for Security Sciences, Riyadh, Kingdom of Saudi Arabia;2. Chimera Translational Research Fraternity Pvt. Ltd., South Extension Part II, 110049, New Delhi, India;3. Arab Society for Forensic Sciences and Forensic Medicine, Naif Arab University for Security Sciences, Saudi Arabia;1. Canary Center at Stanford for Cancer Early Detection, Bio-Acoustic MEMS in Medicine (BAMM) Laboratory, Department of Radiology, Stanford School of Medicine, Stanford University, Palo Alto, CA 94304, USA;2. School of Criminal Justice, Nova Southeastern University, Fort Lauderdale, FL 33314, USA;3. DxNow Inc., Gaithersburg, MD 20879, USA |
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| Abstract: | This investigation was undertaken as an efficient method for isolating sperm DNA from a mixed fluid sample which contains vaginal epithelial cells in a greater amount. The modified method of the two-step differential extraction procedure was found to be suitable for separating sperm DNA and vaginal epithelial cell DNA from the mixed stains. As the first step of digestion, vaginal epithelial cells in the mixed stains were lysed with Proteinase K and SDS, and sperm heads remaining in the lysed solution were collected by centrifugation. As the second step digestion, the sperm heads were lysed with the buffer containing Proteinase K, SDS and DTT as reducing agent. DNA fractions extracted from the two lysed solutions were enriched, one with sperm DNA and the other with vaginal epithelial cell DNA. MCT118(D1S80), ApoB VNTR and HLADQα types of sperm DNA were detected and were confirmed by matching with corresponding male blood DNA. In the case of vaginal secretion mixed with semen of two males, the mixture of MCT118 types of the two males was detected in sperm DNA fraction. |
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