16S V4 of corn sillage tissue - replicate 5
Tissue samples were collected from maize silage at various stages of fermentation. Total DNA was extracted directly from 250 mg of silage biomass using the Magen HiPure Soil DNA Kit following the manufacturers protocol. DNA concentration was measured fluorometrically with a Qubit 4.0 instrument (Thermo Fisher Scientific, USA). V4 region 16S rRNA amplicon libraries were prepared using a two-step PCR, with primers 515F/Pro-mod-805R and dual six-nucleotide indices for multiplexing. Libraries were pooled equimolarly and sequenced on an Illumina MiSeq platform (2  156 bp paired-end reads).