一种改进的方法提取白头鹤粪便DNA  

An Improved Method for DNA Extraction from the Feces of Hooded Crane Grus monacha

作　　者：代艳丽[1] 周立志[1] 

机构地区：[1]安徽大学资源与环境工程学院,安徽大学生物多样性与湿地生态研究所,安徽省生态工程与生物技术重点实验室,合肥,230601

出　　处：《野生动物》2011年第4期203-207,223,共6页

基　　金：国家自然科学基金项目(30470257);安徽大学人才队伍建设项目

摘　　要：现有的粪便DNA提取方法通用性不好,DNA纯度不高不适用于白头鹤等水鸟,本研究探索一种改进的硫氰酸胍裂解法。通过线粒体DNA细胞色素b、控制区基因和微卫星DNA的PCR扩增及线粒体DNA控制区序列的测定,经琼脂糖凝胶电泳检测以及测定序列显示,提取的DNA模板纯度高,效果很好,证实改进方法的可靠性、有效性。为其他濒危水鸟的非损伤取样提供了新的材料和方法,为其保护遗传学的研究提供一定的新技术。DNA extraction from feces has been complicated by two methodological problems：Most methods are not universally applied and the purity of the DNA is often in adequate to water birds.Previously used methods were found unsuitable for water birds such as Hooded Crane（Grus monacha）.In this study we set out to improve the silica-guanidine thiocyanate method for extracting DNA from feces of the Hooded Crane.We isolated and purified DNA using an improved silica-guanidine thiocyanate method.Mitochondrial DNA cytochrome b gene,control region,and microsatellite DNA were amplified successfully from the feces samples,and mitochondrial DNA was sequenced.The electrophoresis and sequence results showed that this method can efficiently extract DNA from Hooded Crane feces.The extracted DNA was of high purity and the method proved reliable and efficient. This is a new approach for noninvasive sampling of Hooded Crane.This type of noninvasive sampling can enhance under-standing of the genetics of highly endangered waterbirds.This method can provide data to evaluate the genetic diversity of Hooded Cranes.This type of data will be important for the development of conservation strategies in future.The procedure is applicable for conservation genetics.

关 键 词：粪便DNA 硫氰酸胍裂解法 线粒体DNA 微卫星 白头鹤 

分 类 号：Q95-33[生物学—动物学]