title: GSM4314331: Animal 7, tumor 1, time 12; Equus caballus; RNA-Seq
library_construction_protocol: Excision of in vivo melanoma tumors for histology and transcriptomics. Total RNA was isolated from cultured cells with Trizol (Ambion, Life Technologies) and purified with an RNAspin Mini kit (GE Healthcare) according to the manufacturer's instructions, with extended treatment with DNase I for 1 h. RNA quality was evaluated with an Agilent 2100 Bioanalyzer RNA Pico assay. RNA was quantified by Quant-iT RiboGreen RNA reagent and Kit (Invitrogen, Life Technologies). Messenger RNA (mRNA) was isolated and used to prepare the complementary DNA (cDNA) libraries following TruSeq RNA Sample Prep Kit V2 (Illumina, San Diego, CA) instructions. Briefly, mRNA was isolated with oligo-dT and purified. Then mRNA was fragmented by heating at 94°C (4 min) in fragmentation buffer. Double-stranded cDNA was synthesized, end-repaired and A-tailed. Sequencing adapters were then ligated to cDNA fragments, according to the manufacturer's protocol. cDNAs fragments were enriched after 15 cycles of PCR amplification. Library quality control was evaluated by the size distribution of the cDNA libraries measured by 2100 Bioanalyzer with DNA1000 assay (Agilent Technologies) and an ABI StepOnePlus Real-Time PCR System was used for quantification of the sample library before sequencing. All 18 cDNA libraries were sequenced on Illumina HiSeq 1500 System, into a rapid paired-end flowcell in 200 cycles of 2x101 bp paired-end strategy.
attributes.tag: GEO Accession
attributes.value: GSM4314331