Jehaes E, Gilissen A, Cassiman J J, Decorte R
Center for Human Genetics, University of Leuven, Belgium.
Forensic Sci Int. 1998 Jun 8;94(1-2):65-71. doi: 10.1016/s0379-0738(98)00052-8.
Mitochondrial DNA (mtDNA) sequencing is a powerful and sensitive method to identify the donor of shed hairs found at a crime scene. Because of the low amounts of DNA in shed hair and the sensitivity of PCR, contaminating cells (e.g. saliva, blood), sometimes present on these hairs, will be co-amplified. This will result in ambiguous sequencing results and might even lead to erroneous exclusions of suspects. We have evaluated a strategy for effectively removing saliva and blood contamination from hair samples. Unambiguous mtDNA results were obtained by incubating the hair samples in a differential lysis buffer (which contains no DTT) prior to DNA extraction. Since the nuclear DNA of the hair root is affected, this procedure should be restricted to hair shaft proportions.
线粒体DNA(mtDNA)测序是一种强大且灵敏的方法,可用于识别在犯罪现场发现的脱落毛发的捐赠者。由于脱落毛发中的DNA含量较低,且PCR具有敏感性,这些毛发上有时存在的污染细胞(如唾液、血液)会被共同扩增。这将导致测序结果不明确,甚至可能导致错误地排除嫌疑人。我们评估了一种有效去除毛发样本中唾液和血液污染的策略。在DNA提取之前,将毛发样本在不含二硫苏糖醇(DTT)的差异裂解缓冲液中孵育,从而获得明确的mtDNA结果。由于发根的核DNA会受到影响,该程序应仅限于毛干部分。