改良高盐低pH方法提取陈旧大麻DNA初探  

A modified method for DNA extraction of stale cannabis sativa samples

作　　者：裴黎[1] 牛慧媛[2] 涂政[2] 彭建雄[2] 马原[2] 从斌[1] 毛泽善[3] 于静娟[4] 

机构地区：[1]河北医科大学,河北石家庄050017 [2]公安部物证鉴定中心,北京100038 [3]新乡医学院,河南新乡453003 [4]中国农业大学,北京100094

出　　处：《中国法医学杂志》2009年第3期145-148,共4页Chinese Journal of Forensic Medicine

基　　金：国家十一五支撑计划基金资助项目2006BAK09B02

摘　　要：目的探讨建立室温保存10年以上大麻干叶及大麻树脂DNA提取方法。方法采用SDS及改良高盐低pH方法,改变提取缓冲液中β-巯基乙醇终浓度,增加用酚、氯仿快速抽提过程,提取新鲜和陈旧大麻(树脂)DNA,应用大麻叶绿体trnL intron引物进行PCR,琼脂糖凝胶电泳法检测扩增产物。结果用高盐低pH方法获得了10年以上大麻干叶及树脂清晰的电泳图谱,其中成功提取了1份23年陈旧大麻的DNA。结论高盐低pH方法操作简便、实用,可望用于陈旧、微量大麻植株的DNA检测,对于涉毒案件中特殊大麻标本的检验具有一定意义。Objective To develop a stable method for extracting the DNA of Cannabis sativa samples （including dry leaves and colophony） which have been kept for more than 10 years in room temperature. Methods Genomic DNA of the fresh and stale cannabis sativa samples was extracted using the SDS method and modified high salt low pH method respectively, and then the trnL gene in the chloroplast of cannabis sativa was amplified and detected. Results Positive result was obtained from all of the samples detected, including a stale sample kept for more than 23 years. Conclusion The improved high salt low pH method was simple and efficient, and can be used to DNA detection of the stale and tittle cannabis sativa plants, which is a important for the identification of cannabis sativa in forensic practice.

关 键 词：法医物证学 陈旧大麻 SDS法 高盐低pH法 DNA 

分 类 号：D919[医药卫生—法医学]