蜘蛛HMW-gDNA的电洗脱纯化提取

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2 mL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（>48.5 kb），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 and Electroelution of Spiders'HMW-gDNA

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.