蜘蛛HMW-gDNA的电洗脱纯化提取  

Extraction and Electroelution of Spiders'HMW-gDNA

作　　者：陈格飞[1] 张云龙[1] 林森珠 李文[1] 孟清[1] 

机构地区：[1]东华大学生物科学与技术研究所,上海201620

出　　处：《Zoological Research》2009年第6期627-632,共6页动物学研究（英文）

基　　金：国家高技术研究发展计划("863"计划)项目(2006AA03Z451);教育部博士点基金项目(08Y10512);东华大学青年教师科研启动基金项目(105-10-0044018);上海市大学生创新活动计划项目(X120704100)

摘　　要：该文研究了蜘蛛大分子量基因组DNA(HMW-gDNA)的提取以及一种高效电洗脱纯化装置的构建。以蜘蛛胸部肌肉组织为原料,通过自改良CTAB法提取蜘蛛HMW-gDNA,利用透析膜和2mL离心管构建一种新的HMW-gDNA快速凝胶回收装置,并对蜘蛛HMW-gDNA进行电洗脱分离回收。结果显示,改良CTAB法可高效提取蜘蛛HMW-gDNA(>48.5kb),且通过透析膜的截留作用,对普通琼脂糖凝胶中目的HMW-gDNA进行快速电洗脱分离,其回收率超过75%,OD260/OD280处于1.8～2.0之间,对HMW-gDNA完整性无影响。综合结果表明,改良CTAB法可用于蜘蛛HMW-gDNA的提取,此电洗脱纯化装置可从普通琼脂糖中高效回收HMW-gDNA,是一种低成本、简捷、高效且实用性强的凝胶回收方法。Extraction of high molecular weight genomic DNA （HMW-gDNA） from spider organs and construction of an electroelution system for obtaining HMW-gDNA efficiently are described. We improved the CTAB method to isolate genomic DNA from spider pectoral muscle tissues, constructed a novel electroelution system with the use of 2-mL centrifuge tube and dialysis membrane, and utilized this self-constructed electroelution system to recover target HMW-gDNA from agarose gels. The HMW-gDNA extracted using the improved CTAB method could be over 48.5 kb in size. By trapping the HMW-gDNA with dialysis membrane, the electroelution system could recover target HMW-gDNA from agarose gels rapidly and without affecting the integrity of HMW-gDNA. The recovery rate was over 75% with the OD260/OD280 ranging from 1.8 to 2.0. The results demonstrate that the improved CTAB method can be used to extract spider HMW-gDNA, and the electroelution system is a general method for recovering target HMW-gDNA from agarose gels with low cost, simplicity, high efficiency and good practicability.

关 键 词：高分子量基因组DNA 蜘蛛 快速分离 电洗脱 

分 类 号：Q781[生物学—分子生物学] Q523