GSM8679232: Bd21-3 roots,  Week 4, Replicate 3  (Root4_3); Brachypodium distachyon; RNA-Seq
Root and shoot tissues were harvested from hydroponically grown B. distachyon Bd21-3 plants at weeks 2 and 4. Total RNA was isolated with a NucleoSpin® RNA Plant Kit (Takara Bio USA) according to the manufacturer's protocol. Three biological replicates were performed using different plants. RNA quality was assessed based on RNA integrity number using an Agilent Bioanalyzer. RNA-seq library construction and sequencing were performed by Novogene. Messenger RNA (mRNA) was isolated from total RNA using poly-T oligo-attached magnetic beads. Following fragmentation, the first strand of cDNA was synthesized using random hexamer primers, and the second strand of cDNA was then synthesized using either dUTP for directional library preparation or dTTP for non-directional library preparation. The libraries were assessed for quantification using Qubit and real-time PCR, and the size distribution was analyzed with a Bioanalyzer. The quantified libraries were pooled and sequenced on Illumina platforms, based on the effective library concentration and desired data yield.