GSM9139260: RNAseq, drought, rep1; Solanum lycopersicum; RNA-Seq
Total RNA isolation: Total RNA was extracted from tomato leaves in three biological replicates (S1, 48h) using Quick-RNA™ Plant Miniprep Kit (Zymo Research, Irvine, CA, USA) according to the manufacturer's protocol. RNA quantity and quality was assessed via microcapillary electrophoresis using a NanoPhotometer® N50 (Implen, Munchen, Germany), and by fragment analysis with an Agilent Bioanalyzer 2100 system using RNA 6000 Nano Kit (Agilent, Santa Clara, CA, USA). Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount.