GSM5769414: S31; Canis lupus; RNA-Seq
Cells were harvested and centrifuged 5 min at 300g. Pellet was suspended in 200 µl PBS and lysed in 550 µl Lysis/binding buffer (Roche) supplemented with 1% DTT (Sigma Aldrich). Cell lysates were stored at -80°C until RNA isolation with the MagNA Pure LC RNA isolation kit-High performance (Roche) on the MagNA Pure LC 2.0 instrument following the manufacturer's protocol. Isolated RNA samples were quantified using UV spectroscopy (Take3 module, Synergy Mx, BioTek, Winooski, USA). RNA integrity was analysed by agarose gel electrophoresis using 2% E-Gel EX pre-stained gels (Thermo Fischer Scientific). Library preparation and RNA sequencing was performed at MAD: Dutch Genomics Service & Support (University of Amsterdam, The Netherlands). Libraries were sequenced using an NextSeq 550 platform and 20 million reads per sample.