GSM9698381: PD-Severe-rep2; Canis lupus familiaris; RNA-Seq
Tissue was homogenized in TRIzol using a bead mill, and then chloroform was added to promote phase separation. The aqueous phase was mixed with an equal volume of 100% ethanol and then RNA was purified using the Zymo Quick RNA kit, including DNAse treatment. RNA concentration was determined with a Nanodrop and integrity assessed on a Fragment Analyzer. Directional RNA-seq libraries were prepared from 500ng total RNA using the NEBNext Directional Ultra II RNA Library Prep Kit for Illumina (New England Biolabs), with initial rRNA depletion using the NEBNext HMR rRNA Depletion reagents.