Rumen bacterial diversity associated to Nitrogen retention efficiency in Feedlot steers
A PCR was employed to amplify the V3 and V4 regions of the 16S ribosomal RNA gene 16S rRNA for bacteria. The expected fragment length of PCR products was verified by agarose gel (1%) electrophoresis, and the amplicon size was estimated by comparison with a 1 kb plus DNA ladder. The PCR fragments were purified using the ZymocleanTM Gel DNA Recovery kit following the manufacturers instructions. Composite samples for sequencing were created by combining equimolar ratios of amplicons. Sequencing was performed using the Illumina MiSeq (Illumina, Inc., NY, USA) with the kit MiSeq Reagent v2 (2x250pb).