GSM5388422: Murine BAT cells_Vehicle rep 1; Mus musculus; RNA-Seq
Cell lysates were collected by administering 175µL per well of RNeasy Lysis Buffer (RLT) (Qiagen, Hilden, Germany) supplemented with 143mM beta-mercaptoethanol (Sigma-Aldrich) and vortexing until cells lifted from plates. Once cells were lifted from plates, 175µL of 70% ethanol was added to each well and lysates were transferred to a High Pure Filter tube from Roche RNA Isolation Kit (Roche Diagnostics, Mannheim, Germany). RNA isolation was done as outlined by manufacturers specifications. Library preparations were conducted using the NEBNext Ultra II Directional RNA Library Prep Kit (NEB, Ipswich, MA, USA) and library fragment size distribution verified using Agilent 2100 Bioanalyser (Agilent, Santa Clara, CA, USA). 