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陈格飞, 张云龙, 林森珠, 李文, 孟 清. 2009: 蜘蛛HMW-gDNA的电洗脱纯化提取. 动物学研究, 30(6): 627-632. DOI: 10.3724/SP.J.1141.2009.06627
引用本文: 陈格飞, 张云龙, 林森珠, 李文, 孟 清. 2009: 蜘蛛HMW-gDNA的电洗脱纯化提取. 动物学研究, 30(6): 627-632. DOI: 10.3724/SP.J.1141.2009.06627
CHEN Ge-fei, ZHANG Yun-long, LIN Sen-zhu, LI Wen, MENG Qing. 2009: Extraction and Electroelution of Spiders’HMW-gDNA. Zoological Research, 30(6): 627-632. DOI: 10.3724/SP.J.1141.2009.06627
Citation: CHEN Ge-fei, ZHANG Yun-long, LIN Sen-zhu, LI Wen, MENG Qing. 2009: Extraction and Electroelution of Spiders’HMW-gDNA. Zoological Research, 30(6): 627-632. DOI: 10.3724/SP.J.1141.2009.06627

蜘蛛HMW-gDNA的电洗脱纯化提取

Extraction and Electroelution of Spiders’HMW-gDNA

  • 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。

     

    Abstract: 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.

     

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