Alvarez Michelle, Juusola Jane, Ballantyne Jack
Graduate Program in Biomolecular Science, University of Central Florida, P.O. Box 162366, Orlando, FL 32816, USA.
Anal Biochem. 2004 Dec 15;335(2):289-98. doi: 10.1016/j.ab.2004.09.002.
RNA analysis is expected to play an increasingly important role in the area of biomolecular forensic analysis. For example, mRNA expression analysis performed on a total RNA sample isolated from a biological stain may be used to identify the nature of the tissue(s) comprising the stain. Many of the physiological stains encountered at crime scenes involve heterogeneous mixtures of different body fluids (e.g., semen and saliva, semen and vaginal secretions). Separate sampling of these mixed stains from different "geographical" locations of the stains to isolate DNA and RNA could result in a misleading estimate of the ratio of the body fluids present and, in extreme cases, even fail to detect one of the contributors. Thus, a prerequisite for the use of mRNA expression profiling in routine forensic analysis is the ability to co-extract DNA and RNA from the same stain. This article describes an optimized method that was specifically developed to co-extract mRNA and DNA from the same physiological stain and that appears to be sufficiently sensitive and robust for routine forensic use.
RNA分析有望在生物分子法医分析领域发挥越来越重要的作用。例如,对从生物污渍中分离出的总RNA样本进行mRNA表达分析,可用于确定构成该污渍的组织的性质。犯罪现场遇到的许多生理污渍都涉及不同体液的异质混合物(例如,精液和唾液、精液和阴道分泌物)。从污渍的不同“地理位置”对这些混合污渍进行单独采样以分离DNA和RNA,可能会导致对存在的体液比例的误导性估计,在极端情况下,甚至可能无法检测到其中一种成分。因此,在常规法医分析中使用mRNA表达谱分析的一个先决条件是能够从同一污渍中共提取DNA和RNA。本文描述了一种经过优化的方法,该方法专门用于从同一生理污渍中共提取mRNA和DNA,并且对于常规法医使用而言似乎具有足够的灵敏度和稳健性。