GSM4985072: WT 24h GP532 rep1; Mus musculus; RNA-Seq
Livers were collected at 30 min or 24 hrs post-treatment and flash frozen. Total and small RNA was extracted using the miRNeasy mini kit (Qiagen) following the manufacturer's recommended protocol. DNAse digestion was performed to remove any residual genomic DNA followed by additional washes prior to elution of high-quality RNA in RNase-free water. The purified RNA was quantitatively assessed using a Qubit Broad Range RNA kit (Thermofisher) and its concentration was determined by Ribogreen fluorescent binding. The RNA was further evaluated qualitatively using RNA Nanotape on the 4200 Tapestation (Agilent technologies), where sizing of the RNA was determined, and a qualitative numerical score (RINe) was assigned. Sequencing libraries were prepared with the RNA HyperPrep Kit with RiboErase (HMR) kit (Roche Sequencing Solutions) from 500 ng total RNA following the manufacturer's recommended protocol. The resulting pool of DNA libraries was then loaded into the appropriate NovaSeq Reagent cartridge for 100 cycle paired end sequencing and sequenced on a NovaSeq6000 following the manufacturer's recommended protocol (Illumina, Inc.).