A comparative study of two extraction methods routinely used for DNA recovery from simulated post coital samples |
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| Institution: | 1. School of Forensic and Investigative Science, University of Central Lancashire, Preston, UK;2. Key Forensic Services, DNA Division, Norwich, UK;1. GENES Ltda, Medellín, Colombia;2. Instituto de Biología, Universidad de Antioquia, Medellín, Colombia;3. Universidad CES, Medellín, Colombia;4. Universidad de Pamplona, Colombia;5. Universidad Pontificia Bolivariana, Medellín, Colombia;1. Laboratorio de Genética Molecular de Cruz Vital – Cruz Roja Ecuatoriana Quito, Ecuador;2. Laboratorio GENES Ltda, Medellin, Colombia;1. Institute of Legal Medicine, Faculty of Medicine, University of Cologne, Cologne, Germany;2. IPATIMUP – Institute of Molecular Pathology and Immunology of the University of Porto, Porto, Portugal;3. Forensic Genetics Unit, Institute of Legal Medicine, University of Santiago de Compostela, Santiago de Compostela, Spain;1. DNAbiotec (Pty) Ltd, Pretoria, South Africa;2. Faculty of Health Sciences, University of Pretoria, Pretoria, South Africa;1. Department of Forensic Genetics, West China School of Basic Science and Forensic Medicine, Sichuan University, Chengdu 610041, Sichuan, PR China;2. Shanghai Key Laboratory of Forensic Medicine, Institute of Forensic Sciences, Ministry of Justice, Shanghai 200063, PR China |
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| Abstract: | In sexual assault cases DNA profiling of spermatozoa can be of critical importance. Most methods use differential extraction of the spermatozoa to separate it from the female component. Here we have compared two commercially available differential extraction methods, the QIAamp® DNA mini kit (Qiagen) and Differex? with the DNA IQ® System (Promega). Simulated postcoital samples were prepared using buccal cells from a female donor and spermatozoa from three male donors. A dilution series ranging from neat semen to a 1:1500 dilution (semen:dH2O) was prepared and mixed with an equal volume of saliva from a female donor. Extraction efficiency was assessed using DNA concentration measured with NanoDrop 2000 and Quantifiler® Human DNA Quantification Kit and the profile count of full, partial and mixed DNA profiles generated using SGM Plus and PowerPlex® ESI 17. Statistical analysis was carried out using Randomisation in R, which is a robust model making no assumption of the distribution of data. Based on the amount of DNA extracted and the types of profiles no significant difference in the performance of the two extraction kits was seen. However, the processing time taken with the Differex? System was about half than that of the QIAamp® DNA mini kit and involved fewer liquid transfers. |
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| Keywords: | Differential extraction Differex? Sexual assault |
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