16S V4 of corn sillage flushing - replicate 2
Flushing samples were collected by incubating 250 mg of silage in 8 mL of 0.1% Tween-20 with constant agitation for 20 minutes to enrich microbial biomass. The suspension was centrifuged at 13,000 rpm for 5 minutes to pellet microbial cells. DNA was extracted from the pelleted biomass using the Magen HiPure Soil DNA Kit with mechanical cell disruption. DNA concentrations were measured with a Qubit 4.0 instrument. V4 region 16S rRNA amplicon libraries were generated using a two-step PCR and dual indexing, pooled in equimolar amounts, and sequenced on an Illumina MiSeq (2  156 bp paired-end reads).