Extraction of PCR-Quality Plant and Microbial DNA from Total Rumen Contents | Zendy

Ranjana Sharma | Zendy; S. Jacob John | Zendy; Dana Damgaard | Zendy; Tim A. McAllister | Zendy

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Extraction of PCR-Quality Plant and Microbial DNA from Total Rumen Contents

Author(s) -

Ranjana Sharma,

S. Jacob John,

Dana Damgaard,

Tim A. McAllister

Publication year - 2003

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/03341st06

Subject(s) - rumen , dna extraction , dna , biology , polymerase chain reaction , extraction (chemistry) , population , bacteria , chromatography , microbiology and biotechnology , biochemistry , chemistry , gene , genetics , fermentation , demography , sociology

DNA from rumen digesta has several diagnostic applications such as studying microbial community dynamics, transgene/DNA stability, and population typing of various rumen bacteria. Several DNA extraction procedures are described in the literature for rumen digesta, which describe the removal of tannins, polysaccharides, and other PCR inhibitors. Some of these protocols are time-consuming and impractical when handling a large number of samples routinely. Here we describe a rapid method for the extraction of PCR-quality plant and microbial DNA from total rumen contents that is based on modifications in the cetyltrimethylammonium bromide procedure followed by cleanup using a Qiagen column. This procedure is highly reproducible and relatively short, once the initial grinding of the samples is performed, and it consistently yields PCR-quality DNA.

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