Barros F, Lareu M V, Salas A, Carracedo A
Institute of Legal Medicine, School of Medicine, Santiago de Compostela, Spain.
Electrophoresis. 1997 Jan;18(1):52-4. doi: 10.1002/elps.1150180110.
A strategy is described for detecting mitochondrial (mt) DNA variation which permits rapid and straightforward screening for forensic purposes. The method is based on the selection of fragments with adequate length for performing single strand conformation polymorphism (SSCP) analysis selecting a set of restriction enzymes (RE) which yield fragments with prefixed lengths. After digestion of mtDNA by the appropriate enzyme or set of enzymes, SSCP analysis is performed in a semiautomatic electrophoretic system using a silver staining detection method. The conformational changes due to single mutations were therefore found not to change the electrophoretic protocol but to change the relative position of the mutations within the fragment. The discrimination power of this method is estimated to be 90% when two restriction enzymes (MspI and HinfI) are used, but it is considerably higher when other enzymes are added.
本文描述了一种用于检测线粒体（mt）DNA变异的策略，该策略可用于法医目的的快速直接筛查。该方法基于选择具有足够长度的片段以进行单链构象多态性（SSCP）分析，选择一组能产生具有预定长度片段的限制性内切酶（RE）。在用适当的一种或一组酶消化mtDNA后，使用银染检测方法在半自动电泳系统中进行SSCP分析。因此发现，单突变引起的构象变化不会改变电泳方案，但会改变片段内突变的相对位置。当使用两种限制性内切酶（MspI和HinfI）时，该方法的鉴别力估计为90%，但添加其他酶时鉴别力会显著更高。
