Illumina HiSeq 2500 paired end sequencing; RNA-seq in the U2OS-AR cell line to identify genes regulated by the androgen receptor
RNA-seq in the U2OS-AR cell line to identify genes regulated by the androgen receptor
RNA was isolated from 1.2 million cells using the RNeasy kit from Qiagen. U2OS-AR cells where grown to approximately 90% confluence according to standard protocols in DMEM medium with 5% FBS. Cells were treated for 24 hours with 5 nM R1881 or 0.1% DMSO as vehicle control. Prior to reverse transcription, poly adenylated RNA was isolated using oligo d(T) beads. Sequencing libraries were prepared using the TruSeq RNA library Prep Kit (Illumina).