GSM5208231: dox 0 ponA 0 MEF; Mus musculus; RNA-Seq
RNA to be utilized for RNAseq experiments was harvested 24 hours post-drug treatment utilizing the Aurum Total RNA Mini kit (Bio-Rad) and DNase treated on column. RNAseq libraries (500 ng, RQN > 9) were prepared using the NEBNext Ultra II Directional RNA Library Kit for Illumina with NEBNext rRNA depletion (Bio-Rad). Protocols were followed as per the manufacturer's guidelines except that 40X adaptor dilutions were utilized, 4X lower concentrations of index primers were used, all bead incubations were performed at room temperature, and 10 cycles of library amplification were performed. Libraries were pooled, and 2 x 76 paired-end sequencing performed using the Illumina NextSeq 500 (Illumina, San Diego, CA, USA).