GSM9627659: Parent tumor samples 74-88-98; Canis lupus familiaris; RNA-Seq
RNA was isolated from flash frozen tumor resections using Trizol reagent (ThermoFisher Scientific, Waltham, MA, USA) following manufacturer's instructions and further purified using the RNeasy kit (Qiagen Valenica CA USA) with the optional on-column DNase step according to the manufacturer's protocols. Indexed, stranded mRNA-seq libraries were prepared from total RNA (1000 ng) using the KAPA Stranded mRNA-Seq Kit (Roche, Indianapolis IN USA) according to the manufacturer's standard protocol for mRNA capture, fragmentation, random-primed first strand synthesis, second strand synthesis with dUTP marking, A-tailing, adaptor ligation, and library amplification. Libraries were pooled and multiplex sequenced on an Illumina NovaSeqX Plus System