GSM3005108: A_Normal_Mucosa; Canis lupus familiaris; RNA-Seq
Normal bladder was removed and bladder mucosa was separated using fine dissection. Bladder tumors were collected via cystoscopy. Tissues were stored directly in trizol and flash frozen. RNA was extracted and purified using RNAeasy (Qiagen).  Ribosomal RNA was removed and libraries constructed using ScriptSeqTM v2 RNA-Seq library preparation kit. Following purification, di-tagged cDNA was amplified by limit-cycle PCR and purified using AMPure XP System (Beckman Coulter) Paired-end 150 bp sequence reads were generated using Illumina HiSeq 4000 platform, to obtain an average of 50 million reads/sample. Raw reads were cleaned for PCR artifacts and adapter trimmed, then aligned to the canine genome (CanFam 3.1 reference genome) using COBWeb to obtain expression levels for annotated genes and isoforms (Strand NGS, v3.1, Build 234566, Strand Life Sciences Pvt Ltd, Bangalore, India)