GSM8148389: 119191_TAATGCGC-ACGTCCTG; Mus musculus; RNA-Seq
RNA was extracted from PBS-perfused, flash-frozen BS/Dien, spinal cord, and cerebellum samples from the left hemisphere. Using the Next Advance Bullet Blender, samples were homogenized in radioimmunoprecipitation assay (RIPA, by Sigma, St. Louis, MO) buffer (750 µL for BS/Dien and 500 µL for cerebellum and spinal cord samples) with added proteinase inhibitor (BioRad, Hercules, CA). A 250 µL aliquot of this lysate was reserved for Western blot analysis and the remaining sample was used for RNA extraction. RNA extraction was conducted using the QIAshredder (Qiagen Hilden, Germany), RNeasy Mini Kit (Qiagen, Hilden, Germany), and DNase I kit (Qiagen, Hilden, Germany), then eluted in RNase-free water, according to manufacturer's instructions. RNA samples were assessed by nanodrop to determine concentration and sample purity. For library preparation, 1 μg of total BS/Dien RNA was submitted to the University of Michigan Sequencing Core for Illumina HiSeq library preparation, quality control, and RNA sequencing. WT and Q84 RNA samples were chosen for RNA sequencing based on RIN number (>7 RIN) and, if applicable, CAG repeat expansion size (>average 72Q) as determined by Laragen, Inc. The University of Michigan Bioinformatics Core generated libraries (Illumina TruSeq) from purified brain RNA samples, completed quality control analysis, and ran 75x75 paired end sequencing on Illumina HiSeq 4000.