title: RNA-Seq of felis catus: adult eye Feline diffuse iris melanoma (FDIM): Late FDIM: Batch A
design: Extraction performed on FFPE sample. First-strand cDNA generated from 50ng of total RNA sample using the SMARTer Stranded Total RNA-Seq Kit v2  Pico Input Mammalian kit (Clontech Laboratories Inc., Mountain View, CA, USA). AMPure XP beads (Beckman Coulter, Brea, CA, USA) used to purify the cDNA library. Depletion of ribosomal cDNA (cDNA fragments originating from highly abundant rRNA molecules) performed using ZapR v2 and R-probes v2 specific to mammalian ribosomal RNA and human mitochondrial rRNA. Uncleaved fragments enriched by PCR before a final library purification using AMPure XP beads. Sequencing (2x100) performed on NextSeq 2000 platform (llumina Inc., San Diego, CA, USA) using NextSeq 2000 P3 Reagents (200 Cycles).