Effect of DNA extraction methods and sampling techniques on the apparent structure of cow and sheep rumen microbial communities.

ABSTRACT: Molecular microbial ecology techniques are widely used to study the composition of the rumen microbiota and to increase understanding of the roles they play. Therefore, sampling and DNA extraction methods that result in adequate yields of microbial DNA that also accurately represents the microbial community are crucial. Fifteen different methods were used to extract DNA from cow and sheep rumen samples. The DNA yield and quality, and its suitability for downstream PCR amplifications varied considerably, depending on the DNA extraction method used. DNA extracts from nine extraction methods that passed these first quality criteria were evaluated further by quantitative PCR enumeration of microbial marker loci. Absolute microbial numbers, determined on the same rumen samples, differed by more

SUBMITTER: Henderson G 

PROVIDER: S-EPMC3770609 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature