Community deoxyribonucleic acids (DNA) extraction
Community DNA was extracted from 0.5 g of rumen
content (fluid and digesta) by the RBB+C method (Yu and
Morrison, 2004). In brief, the RBB+C method employs two
rounds of bead beating in the presence of NaCl and SDS,
followed by sequential ammonium acetate and isopropanal
precipitations. The precipitated nucleic acids were then
treated with RNase A and proteinase K, and the DNA was
purified using columns from QIAgen DNA Mini Stool Kit
(QIAGEN, Valencia, CA), according to manufacturer's
recommendations.