腐败及角化组织的DNA提取及mtDNA(CA)n重复子分型

Extracting of DNA and Genotyping of Mitchondrial CA Dinucleotide Repeats in Decomposed Tissue and Keratinized Tissue

目的建立腐败及角化组织的mtDNA(CA)n重复子分型方法。方法收集腐败肌肉、指(趾)甲及毛干各10份,采用Chelex-100法、含有DTT的Chelex-100处理法及PCR缓冲液处理法三种方法提取DNA,经PCR扩增,PAG电泳分离、银染技术进行mtDNA(CA)n重复子分型。结果10份腐败肌肉用3种方法提取DNA均获得分型成功;10份指(趾)甲及毛干,用Chelex-100法提取DNA,只有1份指甲分型成功,用含有DTT的Chelex-100处理法及PCR缓冲液处理法提取DNA均获得分型成功。结论用本文建立的方法提取DNA适用于腐败及角化组织的mtDNA(CA)n重复子分型检验,在法医学上具有一定的应用价值。

Objective To establish a genotyping method of the mitchondrial （mtDNA） CA dinucleotide repeats in decomposed tissue and keratinized tissue. Methods DNA was extracted from 10 samples gathered respectively from decomposed tissue, fingernail（toenail） and hair stem treated with Chelex-100, Chelex-100 containing dithiothreitol（DTT） and the polymerse chain reaction（PCR） buffer. And genotyping method of the mtDNA CA dinucleotide repeats was also explored by using PCR amplifying, polyacrylamide gel electrophoresis followed by silver staining. Results Ten decomposed tissue samples and keratinized tissue samples treated with Chelex-100 containing DTY or PCR buffer were typed successfully. Only one of the ten keratinized tissues treated with Chelex-100 was a success. Conclusion The method established to extract DNA applies to genotype the mtDNA CA dinucleotide repeats in decomposed tissue and keratinized tissue and is also a useful maker in forensic genetics.