alias: GSM4872999
title: GSM4872999: E5642 - ALS Cre Control; Mus musculus; RNA-Seq
library_construction_protocol: Mice were anesthetized with Ketamine and Xylazine. The spinal column was rapidly removed, and the lumbar spinal cord was retrieved. Lumbar spinal cords were flash frozen and stored at -80˚C until processed. Frozen tissue was kept on dry ice until homogenized in Qiazol reagent (Qiagen Inc, Cat: 79306). Crude homogenate was mixed with chloroform (70µL per 350µL homogenate) to facilitate removal of lipids, and allowed to equilibrate at room temperature for 2 min. Spun at 13,000 x g for 15 min at 4˚C. The aqueous phase containing the RNA was transferred to a new tube. Then the following steps were completed as detailed by the manufacturer's instructions using an RNeasy Mini Kit (Qiagen, Cat: 74104) with on-column DNAse digestion (Qiagen, Cat:79254). RNA concentration was determined via a NanoDrop spectrophotometer (ThermoFisher, Model: ND-1000) and RNA integrity (RIN) was measured using RNA 6000 Pico Kit in a 2100 Bioanalyzer (Agilent). High quality lumbar spinal cord RNA samples, as confirmed by Bioanalyzer, (RINs: 7.9-8.7) free of DNase and RNase were prepared for sequencing using the TruSeqStranded mRNA library prep kit (Illumina, Cat: 20020594) and the NovaSeq 6000 SP Reagent Kit v1.5 (Illumina, Cat: 20028401). High-throughput RNA sequencing (RNA-seq) was completed using an Illumina NovaSeq 6000 sequencer performed by the NYU Grossman School of Medicine Genome Technology Core.
identifiers.namespace: GEO
identifiers.id: GSM4872999
attributes.tag: GEO Accession
attributes.value: GSM4872999