Illumina NovaSeq 6000 paired Sequencing of pair (KAS24117306) (KAE24092866)
Leaves detached from Col plants at 12 DAE were floated on 3 mM MES (pH 5.8) with or without 30 micrometer cis-Pt and incubated at 22 degree C under 16-h light/8-h dark photoperiod for 1 day. The mRNA-seq experiments were performed in two independent biological replicates per treatment (0 and 30 micrometer cis-Pt). Total RNA was isolated from the leaves using QIAzol (QIAGEN, Germany) and its integrity was assessed using Agilent Technologies 2100 Bioanalyzer (Agilent Technologies, USA). Total RNA samples with an RNA integrity number (RIN) greater than 7.8 were used for mRNA-seq analysis. Poly(A) mRNA was isolated from the total RNA (2 microgram) and fragmented using Illumina TruSeq Stranded mRNA LT Sample Prep Kit (Illumina, USA) with poly-T oligo-attached magnetic beads. After the cDNA synthesis from the RNA fragments using Superscript II reverse transcriptase (InvitrogenTM, USA), strand-specific cDNA libraries were generated by adaptor ligation and sequenced by Illumina NovaSeq 6000 system (Illumina, USA), according to the Illumina Directional RNA-Seq protocol.