GSM6209801: 120 minutes Auxin rep 2; Arabidopsis thaliana; RNA-Seq
All RNA extractions were performed using the Arcturus Pico Pure (Thermo Fischer) RNA extraction kit following manufacturer's recommendations. RNA quality was assessed with a Bioanalyser and only samples with a RIN > 6 were used. Before sequencing, an aliquot of RNA (500 ng) from selected samples was used for cDNA synthesis using Superscript Reverse Transcriptase II and the expression of few targets genes (AHP6, PIN1, MP, PID) as well as a control gene (AT1G04850) tested using qPCR. Four biological replicates were initially generated for all samples and three were sequenced (those with highest RINs). All the sequencing reactions were performed at the “Plateforme Transcriptome” located in Unité de Recherche en Génomique Végétale (URGV) at the Université d'Evry Val d'Essonne (now Institute of Plant Sciences Paris-Saclay (IPS2), Université Paris-Sud, Université d'Evry, Université Paris-Saclay). Libraries were prepared using the Agilent RNA6000 Nano kit and sequenced on a HiSeq2000 (Paired End 2x100b). Fastq files of the reads (with adaptors, reads with quality below 20 and reads mapping to the ribosomes removed) were downloaded from the sequencing lab server.