FC1245_AMI_1
RNA from FC1245 cells was isolated using Trizol (Invitrogen) and the RNeasy mini kit with on-column DNase digestion (Qiagen). RNA purity was assessed by Agilent 2100 Bioanalyzer. Sequencing libraries were prepared from 100 ng of total RNA using the TruSeq Stranded Total RNA sample preparation kit (Illumina) according to the manufacturer's protocol. Briefly, mRNA was purified, fragmented and used for first- and second-strand cDNA synthesis followed by adenylation of 3 ends. Samples were ligated to unique adaptors and subjected to PCR amplification. Libraries were then validated using the 2100 BioAnalyzer (Agilent), normalized and pooled for sequencing. RNA-Seq libraries prepared from two replicates for each experimental condition were subjected to single-ended sequencing on the Illumina NovaSeq 6000 using barcoded multiplexing and a 90-bp read length. Image analysis and base calling were done with Illumina software.