RNA-seq of hypoxic and normoxic soybean roots (cultivars: Embrapa 45 and BR 4)

RNA-sequencing of twelve treatments: two cultivars (Embrapa 45 and BR 4), two oxygen conditions [fully aerobic state (normoxy) and hypoxic], and three treatment sampling times (0.5h, 4h, and 28h). For each of twelve treatments, equimolar quantities of purified total RNA from roots of twelve plants [three biological replicates (four plantlets per replicate)] were pooled to result one library. After processing the twelve libraries (poly-A purification, fractionation, cDNA synthesis using random primers, and ligation to bar-coded adapters), fragments of 150–250 pb were isolated and multiplexed, resulting one sequencing library (a pooled of equimolar quantities from twelve initial libraries; each library with a specific barcode for further bioinformatic discrimination). Sequencing library was used to produce clusters for a 1 x 100 bp single end-sequencing run into one lane on a flow cell for sequencing in a Hi-Seq 2000 (Illumina).