GSM7011820: Liver, AAV8-AGXT, 308; Mus musculus; RNA-Seq
Livers were harvested at endpoint (week 24). Total RNA from mouse liver samples was extracted using QIAGEN's RNeasy kit. Samples were quantitated with a Qubit RNA assay (ThermoFisher Scientific) and RNA quality was determined with the Agilent TapeStation RNA assay (Agilent Technologies). Libraries were prepared with the Stranded mRNA Prep, Ligation Kit (Illumina). One ug of RNA was processed for each sample: mRNA was purified and fragmented. cDNA was synthesized, and 3' ends were adenylated. Anchor sequences were ligated to each sample and limited-cycle PCR was performed to amplify and index the libraries. The average library size was determined using an Agilent TapeStation D1000 assay (Agilent Technologies) and libraries were quantitated with qPCR (Bio-rad CFX96 Touch Real-Time PCR, NEB Library Quant Kit for Illumina).  Libraries were normalized to 0.5 nM and pooled.  The library pool was denatured and diluted to approximately 100 pM.  A 1% library of 2.5 pM PhiX was spiked in as an internal control.