plasma 60s_3
Total RNA was extracted from three biological replicates obtained from 6-day-old seedlings (up to 100 mg) using Precellys (Bertin) and lysing kit with 1.4 mm zirconium beads in 0.5 mL tubes. The settings used were 6000 rpm for 30 seconds, followed by a 10 seconds break, and finished with 6000 rpm for 30 seconds, all done at 4C. In-nuPREP Plant RNA kit (Analytic Jena) was used for RNA isolation and quantified with nanodrop (DS-11 Microvolume Spectrophotometer). RNA quality was assessed on a Fragment Analyzer (Agilent Technologies) and all RNAs had a RNA quality number (RQN) above 7.9. Library preparation and RNA-seq was performed at the Lausanne Genomic Technologies Facility, University of Lau-sanne, Switzerland (https://www.unil.ch/gtf). RNA-seq libraries were prepared from 400 ng of total RNA with the Illumina TruSeq Stranded mRNA reagents (Illumina) using a unique dual indexing strategy, and following the official protocol automated on a Sciclone liquid handling robot (PerkinElmer). Libraries were quantified by a flu-orimetric method (QubIT, Life Technologies) and their quality was assessed on a Fragment Analyzer (Agilent Technologies). Cluster generation was performed with 2 nM of an equimolar pool from the re-sulting libraries using the Illumina HiSeq 3000/4000 SR Cluster Kit reagents, then se-quenced on the Illumina HiSeq 4000 SR platform (single end) using HiSeq 3000/4000 SBS Kit reagents for 150 cycles (single end). Sequencing data were demultiplexed using the bcl2fastq2 Conversion Software (version 2.20, Illumina).