3种提取淡水涡虫基因组DNA方法比较  

Comparision of Three Kinds of Genomic DNA Extraction for Freshwater Planarians

作　　者：孙健[1] 张合彩[1] 陈广文[1] 刘德增[2] 

机构地区：[1]河南师范大学生命科学学院,河南新乡453007 [2]黑龙江省科学院自然资源研究所,哈尔滨150031

出　　处：《河南师范大学学报（自然科学版）》2009年第2期168-171,共4页Journal of Henan Normal University(Natural Science Edition)

基　　金：国家自然科学基金(30670247,30170119,30870368);教育部高等学校博士点基金(200804760003);河南省杰出青年科学基金(0312001100);河南省高等学校创新人才培养工程基金(豫教高[2005]126号);河南师范大学博士科研启动基金(0713)

摘　　要：分别用试剂盒法、改良的CTAB法、改良的酚-氯仿抽提法提取活体涡虫和95%酒精固定的涡虫标本的基因组DNA.结果显示:试剂盒法提取的基因组DNA质量最好,主要表现在分子完整,几乎无降解,并且该方法所得DNA的产率也比另外两种方法的产率要高,试剂盒法是最适合涡虫基因组DNA制备的方法.相同起始物质量条件下,3种方法从活体涡虫中提取的DNA量和纯度均高于95%酒精固定的涡虫标本,但试剂盒法提取95%酒精固定半年以上的涡虫标本,所得DNA产率较高、质量较好,可满足PCR扩增等分子生物学实验的要求.This paper deals with genomic DNA extraction from live planarian and ethanol fixed planarian specimen with DNA extraction kit, improved phenol-chloroform extraction and improved CTAB method. The result shows that the quality of genomic DNA by kit method is higher than the other two methods, the DNA molecules are complete and almost not degraded;furthermore, the DNA yields of this method are also the highest. Therefore, the kit method is the most suitable method for planarian genomic DNA extraction. Compared with live planarian and ethanol fixed planarian specimen, the yields and purity of genomic DNA of the former are higher than the latter for these three DNA extraction methods. But the quality of DNA extracted from planarian specimens fixed by 95% ethanol for more than half one year with kit method is better, and it is qualified for PCR amplification and other molecular experiment. This is significant for planarian collection, transportation and preservation.

关 键 词：淡水涡虫 DNA提取 试剂盒法 改良CTAB法 改良酚-氯仿法 

分 类 号：Q959[生物学—动物学]