Extraction of PCR-quality plant and microbial DNA from total rumen contents.
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.
Sharma
Ranjana
R
Agriculture and Agri-Food Canada Research Centre, P.O. Box 3000, Lethbridge, Alberta, T1J 4B1, Canada.
John
S Jacob
SJ
Damgaard
M
M
McAllister
Tim A
TA
D000465:Algorithms; D000818:Animals; D004269:DNA, Bacterial / Q000302:isolation & purification*; D018744:DNA, Plant / Q000302:isolation & purification*; D005766:Gastrointestinal Contents; D016133:Polymerase Chain Reaction / Q000379:methods*; D012417:Rumen / Q000378:metabolism* / Q000382:microbiology*; D012756:Sheep
D004269:DNA, Bacterial; D018744:DNA, Plant