title: NextSeq 500 paired end sequencing; RNA-seq of Solanum Lycopersicum and Solanum Pennellii parents and crosses up to the F4 generation
design: RNA-seq of Solanum Lycopersicum and Solanum Pennellii parents and crosses up to the F4 generation
library_construction_protocol: 1-4cm young leaves were collected in less than 2h for all plants at the same time of the day using liquid N2 greenhouse grown plants, Approx 24C day, 18 night. 16/8h day/night Trizol isolation. Total RNA isolation was performed with the Direct-zol RNA Miniprep (Zymo Research) and Trizol reagent (Invitrogen) according to the manufacturer's instructions RNA samples were pre-processed to remove Ribosomal content with Ribo-Zero™ Magnetic Kit*(Plant Leaf) (epicentre: Cat. No. MRZPL1224 – 24 Reactions). Subsequent samples were processed using ScriptSeq™ v2 RNA-Seq Library Preparation Kit (epicentre: SSV21124 – 24 Reactions).  RNA was treated with Turbo DNase free kit (Thermo Fisher).