RNA Extraction and Sequencing for Plants

Three biological replicates, corresponding to a pool of leaves from four plants for the distal sampling, and to individual stems (−1 cm) for the local sampling, were used for RNA extraction. Total RNA was isolated using a CTAB-based extraction protocol followed by a DNase treatment as described in Blanco-Ulate et al. (2013b (link)). RNA concentration and purity were measured using a Quibit fluorometer (Thermo Scientific) and a NanoDrop 2000c Spectrophotometer (Thermo Scientific), respectively. Libraries were prepared using the Illumina TruSeq RNA sample preparation kit v.2 (Illumina, CA, USA). Final libraries were evaluated for quantity and quality using the High Sensitivity DNA kit on a Bioanalyzer 2100 (Agilent Technologies, CA).