Comparison of two silica-based extraction methods for DNA isolation from bones |
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Affiliation: | 1. Department of Legal Medicine, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba City, Chiba Prefecture 260-8670, Japan;2. Department of Forensic Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan;1. Netherlands Forensic Institute, The Hague, The Netherlands;2. Dept. of Pathology, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands;3. Barge’s Anthropologica, Dept. of Anatomy, Embryology and Physiology, Leiden University Medical Centre, Leiden, The Netherlands;4. Section of Forensic Medicine, Jakarta Metropolitan Police, Jakarta, Indonesia;5. DNA laboratory, Police Medicine and Health Services Centre, Indonesia National Police, Jakarta, Indonesia;6. Faculty of Science IBED, University of Amsterdam, Amsterdam, The Netherlands;1. Forensic Science Program, Division of Health and Applied Sciences, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, Thailand;2. Forensic Science Innovation and Service Center, Prince of Songkla University, Hat Yai, Songkhla, Thailand;3. Division of Pathology, Faculty of Medicine, Prince of Songkla University, Hat Yai, Songkhla, Thailand;4. Anatomy Program, Division of Health and Applied Sciences, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, Thailand;1. State Key Laboratory of Pollution Control and Resource Reuse, School of the Environment, Nanjing University, Nanjing 210023, China;2. School of Environmental Science and Engineering, Nanjing Tech University, Nanjing 211816, China;3. Department of Civil and Environmental Engineering, Rice University, Houston 77005, USA |
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Abstract: | One of the most demanding DNA extractions is from bones and teeth due to the robustness of the material and the relatively low DNA content. The greatest challenge is due to the manifold nature of the material, which is defined by various factors, including age, storage, environmental conditions, and contamination with inhibitors. However, most published protocols do not distinguish between different types or qualities of bone material, but are described as being generally applicable. Our laboratory works with two different extraction methods based on silica membranes or the use of silica beads. We compared the amplification success of the two methods from bone samples with different qualities and in the presence of inhibitors. We found that the DNA extraction using the silica membrane method results an in higher DNA yield but also in a higher risk of co-extracting impurities, which can act as inhibitors. In contrast the silica beads method shows decreased co-extraction of inhibitors but also less DNA yield. Related to our own experiences it has to be considered that each bone material should be reviewed independently regarding the analysis and extraction method. Therefore, the most ambitious task is determining the quality of the bone material, which requires substantial experience. |
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Keywords: | DNA extraction Bones Silica membrane Silica beads Sample quality |
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