title: GSM8686592: FBD9; Canis lupus familiaris; RNA-Seq
library_name: GSM8686592
library_construction_protocol: RNA extraction and RNA sequencing (RNASeq) were performed at the University of Minnesota Genomics Center (UMGC). RNA was extracted and purified from scrolls using the PureLinkTM FFPE Total RNA Isolation Kit (Invitrogen, Carlsbad, CA). After elution of RNA, DNAse I digestion was performed to yield DNA-free total RNA. Total eukaryotic RNA isolates were quantified using a fluorimetric RiboGreen assay (Life Technologies, Carlsbad, CA). Total RNA integrity was assessed using capillary electrophoresis on the Agilent BioAnalyzer 2100 (Agilent, Santa Clara, CA), generating an RNA Integrity Number (RIN). A RIN cutoff of 2 was used for samples to be acceptable. Samples that passed quality control underwent library preparation using the SMARTer Stranded Total RNA-Seq Kit v2, Pico Input Mammalian (Takara Bio, Mountain View, CA); this kit removes ribosomal RNA. RNA sequencing was performed on the NextSeq 2K, which produces 2 x 50 bp paired end reads (Illumina, San Diego, CA) at a targeted depth of ≥40 million reads per sample.