RNA-seq of olea Europaea var. Empeltre extra virgin olive oil
Library preparation and sequencing were carried out at Fundacin Parque Cientfico de Madrid(Madrid, Spain). Briefly, total RNA was used as input for library preparation, using the NEBNext Multiplex Small RNA Library Prep Set for Illumina (New England BioLabs Ipswich, MA, USA) following the manufacturers instructions. Equimolecular pools of small RNA regions (< 200 nt) werepurified by polyacrylamide gel electrophoresis. The library pool was denatured and seeded on a NextSeq v2.5 flowcell (Illumina, San Diego, CA, USA) and sequenced using a NextSeq 500 High Output kit v2.5 (Illumina) in a 1x75 single-read sequencing run on a NextSeq 500 sequencer (Illumina). In order to generate clean data, the initial sequences were purged from low-quality reads, repeated and adaptor sequences. Bowtie2 was used for alignment against the reference miRNA database from Olea europea L . Feature counting was carried out using HTSeq-count. Before differential expression analysis, the obtained count matrix was normalized using the TMM method available within the edgeR library from the R coding environment.