GSM3544268: CRB_ASO_52w_3; Mus musculus; RNA-Seq
For in vivo mouse samples, total RNA and DNA was extracted from the liver and the right hemisphere's hippocampus, cortex, and cerebellum with AllPrep® DNA/RNA/miRNA kit (Qiagen). Total RNA-seq libraries were prepared using the TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero Gold (Illumina – San Diego, CA) according to manufacturer's instructions. 1µg of RNA was used as starting material and 12 PCR cycles were used to minimize bias. Library size was observed on an Agilent 2100 Bioanalyzer and quantity was determined using qPCR (KAPA Biosystems - Wilmington, MA). Libraries were sequences on an Illumina HiSeq 2500 using a 50-cycle rapid run kit or an Illumina NextSeq instrument using a 75-cycle high throughput kit. Reads were aligned to the GRCm38.p3 mouse genome and transcriptome from iGenomes using TopHat and differential expression tests were performed using featureCounts and edgeR with standard settings.