藏酋猴毛干DNA的3种提取方法  

Comparison of Methods for DNA Extraction from Hair Shaft of Macaca thibetana

作　　者：徐怀亮[1] 汪宴廷[1] 姚永芳[1] 黄晓峰[1] 倪庆永[1] 程安春[1] 潘阳[2] 

机构地区：[1]四川农业大学,雅安625014 [2]昆明动物园

出　　处：《东北林业大学学报》2010年第2期58-61,共4页Journal of Northeast Forestry University

基　　金：国家自然科学基金资助项目(30970383);中国博士后科学基金项目(2005037803);四川省教育厅自然科学基金重点项目(08ZA076);四川农业大学"双支计划"项目(01570104)

摘　　要：为寻求一种从藏酋猴毛干中提取总DNA的有效方法,从一藏酋猴个体背部剪取数量不等的毛干样品(不含毛囊),经蛋白酶K消化后,分别采用高盐沉淀抽提法、酚氯仿抽提法、纳米磁珠抽提法分离获得用于PCR扩增的模板DNA,通过紫外分光光度计测定OD值并计算出质量浓度,同时进行琼脂糖凝胶电泳和PCR扩增检测,分析3种方法提取藏酋猴毛干DNA的质量差异。每处理设置10个重复。结果表明,纳米磁珠法提取藏酋猴毛干中DNA是一种快捷、简便、经济、高效、安全的有效方法;进行毛发取样时以5～10根较为合适。In order to find an effective method to extract DNA from the hair shaft of Macaca thibetana,different numbers of hair shafts without bottom follicle were sampled from dorsal area of a tibetan macaque,which were digested with SDS-proteinase K and extracted by high salt method,phenol-chloroform method and magnetic bead method.As a result,total genomic DNAs to be used as templates for PCR amplification were obtained.Ten repetitions per sample were set in the present study.The extraction results of the three methods were detected through agarose gel electrophoresis and PCR amplification,and optical density value was determined with an ultraviolet spectrophotometer and the content of DNA was calculated.The analysis result indicates that the magnetic bead extraction method is a quick,simple,economical and efficient method to extract DNA from the hair shaft of tibetan macaque.In addition,5~10 hair shaft samples should be used in the experiment.

关 键 词：藏酋猴 毛干 DNA 高盐沉淀抽提法 酚氯仿抽提法 磁珠抽提法 

分 类 号：Q781[生物学—分子生物学]