GSM5350278: Case20; Canis lupus familiaris; RNA-Seq
Cryopreserved tissue was homogenized in Trizol on a bead mill using 2.8mm ceramic beads. Total RNA was isolated from the Trizol lysate, with an extra chloroform extraction to remove residual phenol and addition of glyco-blue as a carrier to promote RNA precipitation. RNA concentration was determined with a Nanodrop and integrity assessed on a Fragment Analyzer. Directional RNA-seq libraries were prepared from 250ng total RNA using the NEBNext Directional Ultra II RNA Library Prep Kit for Illumina (New England Biolabs), with initial polyA+ enrichment.