狗獾粪便DNA提取方法的初步探讨  

A preliminary study on the DNA extraction method from faeces of badger (Meles meles)

作　　者：刘宇庆[1,2] 吕慎金[3] 殷宝法[1] 范红梅[1] 刘燕 魏万红[1] 

机构地区：[1]扬州大学生物科学与技术学院,扬州225009 [2]扬州市邗江区农林局,扬州225002 [3]临沂师范学院农林学院,临沂276000

出　　处：《生态科学》2008年第6期468-473,共6页Ecological Science

基　　金：国家自然科学基金(30570289)

摘　　要：在京杭大运河扬州段堤坝上采集狗獾的新鲜粪便,采用预处理-酚/氯仿抽提法、NaCl改良法、异硫氰酸胍裂解法、淀粉吸附法及QIAamp试剂盒法5种方法对粪便样品中狗獾DNA进行提取,探讨狗獾粪便样品中DNA提取方法和优化条件。结果表明,在5种提取方法中,淀粉吸附法的效果明显优于其它4种方法。采用粪便裂解液快速裂解细胞后,加入淀粉去除其中的大量PCR抑制物,然后用蛋白酶K裂解、酚/氯仿/异戊醇抽提,最后使用UNIQ-10柱纯化粪便DNA,对线粒体控制区和微卫星位点的PCR扩增反应及测序结果证实该方法的可行性。以上结果表明,通过该方法获得的粪便DNA能够用于更深入的分子遗传学等学科的研究。The fresh feces of badger （Meles meles） were collected from the dam of Jing-Hang Canal near Yangzhou, and five DNA extraction methods including amylum-subsiding method, preprocess-organicsolvent extraction method, NaCI modified method, GuSCN-SiO2 method and QIAamp DNA Stool Mini Kit method were used to extract the DNA from them. The results showed that the highest amplification success rate was in the amylum-subsiding method. In this study, fecal sample was lysed rapidly with digested solution, then DNA was extracted using chloroform/phenol after incubation with proteinase K, following by purification to remove numerous potential PCR inhibitors. The mitochondrial DNA control region and 5 microsatellite loci were amplified, showing that the mitochondrial DNA control region sequenced successfully. Our results suggested that DNA extraction based on this method was reliable.

关 键 词：狗獾 粪便DNA 提取 PCR扩增 

分 类 号：Q523.8[生物学—生物化学]