RNA-Seq of Canis lupus familiaris with canine cutaneous histiocytoma group 1: male skin
Quality of total RNA was assessed with the Fragment Analyzer 5200 (Agilent) using the High Sensitivity RNA quantification kit and Fragment Analyzer Controller Software (Agilent v 3.1.0.12). The DV200 score for FFPE-derived total RNA was calculated. Random primed library preparation was started with 100-150 ng of total RNA using the Watchmaker RNA library prep kit with Polaris depletion (Watchmaker Genomics) according to the instructions of the manufacturer. Because the RNAs were partially degraded, we used the Watchmaker FFPE RNA workflow. The barcoded libraries were purified and quantified using Qubit Fluorometric Quantification (ThermoFisher Scientific). Size distribution of the library DNA was analyzed using the Fragment Analyzer 5200 (Agilent). Sequencing of 2x150 bp was performed with an Illumina NovaSeq sequencer (Illumina, San Diego, CA, USA) at the sequencing core facility of the Faculty of Medicine (University Leipzig) according to the instructions of the manufacturer.