title: GSM7051748: H35.RTX.T9.C3; Sorghum bicolor; RNA-Seq
library_name: GSM7051748
library_construction_protocol: Between ~2 and ~40 ug of total RNA was extracted from three biological replicates of sorghum samples for each genotype and treatment (control, heat, cold stress) using GeneJET Plant RNA Extraction Kit (Thermofisher K0801) followed by DNAse I treatment (Thermofisher EN0521). Total mRNAs were isolated using biotinylated oligo(dT) and streptavidin magnetic beads (New England Biolabs) as previously described (Wang et al., 2011, 10.1371/journal.pone.0026426) Purified mRNAs were used for library preparation as previously described with the following modifications. In the final enrichment step, indexed adapter enrichment primers were used (Townsley et al., 2015, 10.3389/fpls.2015.00366) and 12 cycles were performed to amplify the libraries. The libraries were quantified using a Qubit 2.0 Fluorescence Reader (Thermo Fisher Scientific) and quality was verified using a Bioanalyzer 2100 (Agilent Genomics). Final libraries were multiplexed and sequenced on the NextSeq 500 (Illumina) at the UC Riverside (UCR) Institute for Integrated Genome Biology (IIGB) Genomics Core facility to obtain 75 nt single-end reads.