alias: GSM1564239
title: GSM1564239: type I IFN treated TO cells; Salmo salar; RNA-Seq
library_construction_protocol: Total RNA was isolated using the RNeasy mini Kit (Qiagen, Hilden, Germany) with on-column DNase treatment according to the manufacturer’s instructions. Equal quantities of total RNA from triplicates of the type I IFN treated, SAV-3 infected and mock-TO-cells were mixed to prepare the pooled RNA sample for RNA-Seq. Total RNA samples were treated with DNase I to degrade any possible DNA contamination. Then the mRNA was enriched using oligo(dT) magnetic beads. The mRNA was fragmented into short fragments (about 200 bp) by mixing with the fragmentation buffer. Thereafter, the first strand of cDNA was synthesized using random hexamer-primer. A buffer containing dNTPs, RNase H and DNA polymerase I was added to synthesize the second strand. The double strand cDNA was purified with magnetic beads. End reparation and 3'-end single nucleotide A (adenine) addition was then performed. Finally, sequencing adaptors were ligated to the fragments and the fragments were enriched by PCR amplification. During quality check (QC step), Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-Time PCR System (Biorad.com) were used to qualify and quantify the sample library. Thereafter, library products were ready for RNA-sequencing using Illumina HiSeqTM 2000, BGI-Hong Kong.
identifiers.namespace: GEO
identifiers.id: GSM1564239
attributes.tag: GEO Accession
attributes.value: GSM1564239