Improved protocol for high-quality co-extraction of DNA and RNA from rumen digesta.
We report an improved method for total nucleic acids extraction from rumen content samples. The method employs bead beating, and phenol-chloroform extraction followed by saline-alcohol precipitation. Total nucleic acids and RNA yield and purity were assessed by spectrophotometric measurements; RNA integrity was estimated using Agilent RNA 6000 Nano Kit on an Agilent 2100 Bioanalyzer. The method provided total nucleic acids and RNA extracts of good quantity and quality. The extraction is not time consuming and it is valuable for ecological studies of rumen microbial community structure and gene expression.
Popova
M
M
INRA Clermont-Ferrand-Theix, 63122, Saint Gènes Champanelle, France.
Martin
C
C
Morgavi
D P
DP
D000818:Animals; D004247:DNA / Q000302:isolation & purification*; D023281:Genomics / Q000379:methods*; D008828:Microbiological Techniques / Q000379:methods*; D008967:Molecular Biology / Q000379:methods*; D012313:RNA / Q000302:isolation & purification*; D012417:Rumen / Q000382:microbiology*
D012313:RNA; D004247:DNA