title: GSM6283065: Non-cold stored 3; Fragaria vesca; RNA-Seq
library_construction_protocol: The ripe Mara des Bois strawberries (Fragaria vesca) immediately after harvest (2 hours) were used as the control (AH). Those placed directly in a chamber at 20 ºC and 80 % RH for two days were the non-cold stored (NCS), while the strawberries transferred at 20ºC for 1 day following the 7-day storage at 1ºC (conditions employed to simulate a time in transit) were used to analyse the air-cold stored fruit (ACS) or CO2 -cold stored fruit (CCS). CCS were pretreated for two days with a gas mixture containing 18 % CO2 + 18 % O2 + 64 % N2 and then air-ventilated for another five days (CCS). The total RNA of three biological replicate samples was extracted from 0.5 g of fruit powder in accordance with Yu et al., (2012). Each biological replicate sample contained a fruit mixture of at least 50 strawberries. A TURBO DNase™ enzyme (Ambion, Austin, Tex.) treatment was used to degrade the contaminant DNA. RNA quantity and purity were measured with the NanoDrop ND-1000 spectrophotometer (Thermo Scientific). RNA integrity was determined by the RNA integrity number (RIN), using a 2100 Bioanalyzer (Agilent). The mRNA from each biological replica was employed to prepare the RNA-Seq libraries with the reagents provided in the Illumina®TruSeq ™ Stranded Total RNA kit protocol (Illumina Inc., San Diego, CA). The size and quality of the libraries were quality controlled in Agilent DNA 2100 Bioanalyzer assay (Agilent). RNA-seq: Each library was sequenced using Illumina HiSeq™ 2500 (Illumina Inc.), in paired-end mode with a read length of 2x50 bp, generating minimally 39 million paired-end reads per replica and passing filter for each RNA-Seq library in a fraction of a sequencing lane of the sequencer following the manufacturer's protocol (max. 200 M/lane).