摘要
针对cyt b基因和ND6基因序列,设计两对引物,以14种常见动物(包括人)生物性检材为对象进行PCR扩增,产物经聚丙烯酰胺凝胶电泳(PAGE)、硝酸银染色进行分析。结果证明,两条电泳条带者为人样本,分别是cyt b基因片段(358 bp)和ND6基因片段(181 bp);一条电泳条带者为动物来源,是358 bp cyt b基因片段。在37.5μL PCR反应体系中最小检出模板量为0.25 pg基因组DNA。检材在4℃、室温和37℃温度下,经过4个月后均可获得正确的种属区分。高度潮湿环境中放置50 d的检材、形成于水泥地面、墙面、泥土等基质上的血痕,本方法可得到正确区分。由此建立的种属鉴定的线粒体基因复合扩增体系,其检测片段短、灵敏度高、操作简易,适合法医学上微量、降解、陈旧检材的种属判定。
Two pairs of primers for the cyt b gene and ND6 gene sequences were designed and biological sample of 14 species of animals（including humans） PCR were amplified,and the products were analyzed by poly-acrylamide gel electrophoresis（PAGE） and stained with silver nitrate.The results demonstrate that the two electrophoretic bands of the cyt b gene fragment（358 bp） and ND6 gene fragment（181 bp） are human samples;and the one electrophoresis strip of 358 bp Cytb gene fragments is animal sources.In 37.5 μl PCR reaction system,the minimum detectable template concentration is 0.25 pg genomic DNA.Samples can be distinguished correctly at 4°C,room temperature and 37 °C,after four months later.Samples placed in a high humidity environment for 50 days and bloodstains formed in the concrete floor or on the walls,and soil matrix,can be distinguished correctly by the methods.The species identification method of the mitochondrial genome multiplex PCR system with short detected fragments are high sensitivity and easy to operate and suitable to determine the trace or degradation or obsolete samples of forensic.
出处
《盐城工学院学报（自然科学版）》
CAS
2012年第1期19-24,共6页
Journal of Yancheng Institute of Technology:Natural Science Edition
基金
国家自然科学基金项目(30872917
81072510)
