title: RNA-Seq of canis lupus familiaris: soild tumor biopsy from oral melanoma-affected dogs
design: Strand-specific RNA-seq libraries were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina (New England BioLabs, Hitchin, UK) as per the manufacturers specifications. In brief, using 0.3 g of total RNA as input, poly(A) mRNA was enriched using the NEB magnetic mRNA isolation module, then fragmented and reverse-transcribed to generate cDNA. An Illumina-specific adaptor was sequentially ligated to the 3 end of the cDNA fragments, purified using the AMPure XP beads (Beckman Coulter, Brea, CA, USA), and finally PCR-amplified (14 cycles) using an appropriate indexing primer that allows further samples multiplexing. The PCR-amplified libraries were purified with the AMPure XP beads (Beckman Coulter, Brea, CA, USA) and then assessed for their quality and fragments distribution using the 2100 Bioanalyzer DNA 1000 assay (Agilent Technologies, Santa Clara, CA, USA).