GSM8486443: etha_24h_0µM_4; Vitis vinifera; RNA-Seq
Liquid nitrogen frozen callus samples were first homogenised to powder. Total RNA from individual calli was extracted using 160 mg of frozen powder with the Promega ReliaPrep™ RNA Tissue Miniprep System kit (Promega, France, Ref. Z6112) following the Promega - ReliaPrep™ RNA Tissue Miniprep System protocol, optimized for RNA extractions of grapevine cell cultures. Then, RNAs underwent DNAse treatment with the Invitrogen™ ezDNase™ Enzyme kit to remove genomic DNA contamination from RNA preparations, following the ezDNase™ Enzyme kit protocol. For each sample, quantity and purity were first assessed by measuring the optical density at 260 nm and 280 nm and with a 1% (w/v) gel electrophoresis. Then, for each sample, RNA Integrity Number (RIN) was measured using an Agilent 2100 Bioanalyzer System microfluidic on-chip capillary electrophoresis device, following the RNA 6000 Nano kit protocol (Agilent Technologies, Germany). Samples with a RIN > 8.8 were selected and then aliquoted to obtain 20 μL RNA samples at 50 ng/μL. mRNAs were sequenced by Illumina NovaSeq 6000 sequencing system with 2 × 150 bp paired-end sequences.The RNA libraries were constructed following the manufacturer's protocol.