Cartozzo Claire, Singh Baneshwar, Boone Edward, Simmons Tal
Integrative Life Sciences Program, Virginia Commonwealth University, Richmond, VA, 23284.
Department of Forensic Sciences, Virginia Commonwealth University, Richmond, VA, 23284.
J Forensic Sci. 2018 Nov;63(6):1830-1835. doi: 10.1111/1556-4029.13792. Epub 2018 Apr 12.
When deaths occur in water, soft tissue decomposes after a temperature-dependent period, making DNA identification dependent on bone. This study examined the effects of water on bone DNA purity and quantity, and determined the best of three extraction methods for isolating DNA. The organic phenol-chloroform method consistently extracted DNA approximating the accepted 260/280 purity value (~1.8); ChargeSwitch® gDNA Plant Kit and DNeasy Blood and Tissue Kit produced fair and unacceptable values, respectively. The purity value for humerus and rib samples was consistent across accumulated degree days (ADD). Significant differences in quantification among extraction methods and between bone types were identified. Ribs and ChargeSwitch® gDNA Plant Kit samples produced the lowest mean Ct values of the bone types and the extraction methods, respectively. Therefore, this study proposes that magnetic bead technology extraction methods and ribs be considered when processing bones that have been submerged in water for any length of time.
当在水中发生死亡事件时,软组织会在一段取决于温度的时间后分解,使得DNA鉴定依赖于骨骼。本研究考察了水对骨骼DNA纯度和数量的影响,并确定了三种DNA提取方法中最佳的一种。有机酚 - 氯仿法始终能提取出接近公认的260/280纯度值(约1.8)的DNA;ChargeSwitch® gDNA植物试剂盒和DNeasy血液和组织试剂盒分别产生了一般和不可接受的值。肱骨和肋骨样本的纯度值在累积度日(ADD)方面是一致的。提取方法之间以及骨骼类型之间在定量方面存在显著差异。肋骨样本和ChargeSwitch® gDNA植物试剂盒样本分别在骨骼类型和提取方法中产生了最低的平均Ct值。因此,本研究建议,在处理任何时长浸泡在水中的骨骼时,应考虑磁珠技术提取方法和肋骨。