GSM9847596: TS-3; Oncorhynchus mykiss; RNA-Seq
Total RNA was extracted from rainbow trout spleen tissues or hepatocytes using Trizol reagent according to the manufacturer's instructions. RNA concentration and purity were assessed using NanoDrop, and RNA integrity was evaluated by agarose gel electrophoresis. RNA samples meeting quality requirements were used for long RNA-seq, small RNA-seq or mRNA-seq library construction. For spleen long RNA-seq, ribosomal RNA was removed from total RNA, and strand-specific long RNA-seq libraries were constructed for mRNA, lncRNA and circRNA expression profiling. These libraries were sequenced on an Illumina HiSeq 4000 platform using paired-end 150 bp reads. For spleen small RNA-seq, small RNAs of approximately 18-30 nt were enriched, ligated to adapters, reverse transcribed and PCR amplified; libraries were sequenced on an Illumina HiSeq Xten platform using single-end 50 bp reads. For hepatocyte mRNA-seq, messenger RNA was enriched from total RNA using poly(A) selection, fragmented, reverse transcribed and amplified to generate libraries, which were sequenced on an Illumina NovaSeq 6000 platform using paired-end 150 bp reads.