title: GSM7329468: YoGI_310_after_heat_stress [A_29]; Triticum aestivum; ssRNA-seq
library_construction_protocol: For each one of the 13 wheat accessions, 2 cm of leaf tissue was taken at the 3-leaf stage and after the 14 days of heat stress. Total RNA was extracted from <100 mg of individual leaf tissue samples using the E.Z.N.A Plant RNA Kit (Omega Bio-Tek, ) including a DNase treatment, according to the manufacturer's protocol. RNA concentration was quantified by both NanoDrop ND-1000 Spectrophotometer, and Qubit 4 Fluorometer and the quality was assessed by Agilent Technology 2100 Bioanalyzer. Samples with RNA Integrity Number (RIN) values >7 were used for the sequencing. Replicates were pooled into 1 sample per accession, per group, at equimolar proportions. The library preparation and sequencing were performed by Novogene UK. The library was obtained using TruSeq Stranded mRNA (Illumina) . The mRNA was purified using poly-T oligo attached magnetic beads, fragmented and the first strand cDNA synthesized using random hexamer primers, whilst the second strand cDNA using dUTP for directional library. End-repair, A-tailing, adapter ligation, size selection,USER enzyme digestion to remove the UTP-containing second strand cDNA, PCR amplification and purification TruSeq stranded mRNA run in the Illumina Novaseq 6000 platform (Illumina, CA, USA) with an 150bp paired-end sequencing strategy