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Zoological Research
Kunming Institute of Zoology, Chinese Academy of Sciences
ISSN: 2095-8137
Vol. 30, No. 6, 2009, pp. 627-632
Bioline Code: zr09095
Full paper language: Chinese
Document type: Research Article
Document available free of charge

Zoological Research, Vol. 30, No. 6, 2009, pp. 627-632

 en Extraction and Electroelution of Spiders’ HMW-gDNA
Chen, Ge-fei; Zhang, Yun-long; Lin, Sen-zhu; Li, Wen & Meng, Qing

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.

Keywords
HMW-gDNA; Spider; Rapid isolation; Electroelution

 
© Copyright 2009 Kunming Institute of Zoology, the Chinese Academy of Sciences
Alternative site location: http://www.zoores.ac.cn/

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