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目的利用随机扩增多态性DNA和简单序列重复区间扩增分子标记检测大麻遗传多样性,并探讨其在法医学中的应用价值。方法收集中国4省6个地区的100株大麻叶子样品,采用CTAB法提取基因组DNA,设计选择11个RAPD引物和13个ISSR引物,采用6%中性聚丙烯酰胺凝胶电泳-硝酸银染色法进行检测,根据出现的条带数目和片段大小等分析大麻的多样性。结果 11条RAPD引物扩增出的片段在200bp以上共52条,其中具有多态性的27条;ISSR引物扩增出126条,其中具有多态性的73条;多态性条带比率分别为51.9%和57.9%,其差异不具有统计学意义(P>0.05)。结论 RAPD和ISSR两种方法均可用于大麻遗传多样性分析,对检测毒品原植物的种类和来源地具有一定的应用前景。
Objective To detect the genetic diversity of cannabis by using random amplified polymorphic DNA (DNA) and simple sequence repeat interval amplification (PCR-RFLP) markers, and to explore its value in forensic medicine. Methods 100 cannabis leaves samples were collected from 6 provinces in 4 provinces in China. Genomic DNA was extracted by CTAB method. Eleven RAPD primers and 13 ISSR primers were designed and synthesized. Six kinds of neutral polyacrylamide gel electrophoresis - silver nitrate staining The tests were conducted to analyze the diversity of cannabis based on the number of bands present and the size of the fragments. Results The amplified fragments of 11 RAPD primers were 52 above 200bp in total, of which 27 were polymorphic; 126 were amplified by ISSR primers, of which 73 were polymorphic; the polymorphic bands were 51.9% and 57.9% respectively, the difference was not statistically significant (P> 0.05). Conclusion Both RAPD and ISSR methods can be used to analyze the genetic diversity of cannabis, which has certain application prospects for detecting the species and origin of the original drug.