GSM8070821: 7940B_DMSO_8hr_1; Mus musculus; RNA-Seq
Total RNA was isolated using the miRNeasy kit (Qiagen). RNA-seq libraries were prepared using 800 ng of total RNA. Ribosomal RNA were removed by enzymatic digestion of the specific probe-bound duplex rRNA, and then fragmented to around 200-300bp with heat in fragmentation buffer (KAPA RNA Hyper+RiboErase HMR, Roche). Double-stranded cDNA was then synthesized by reverse transcription and underwent end-repair and ligation using New England Biolabs (NEB) adapters. Final library preparation was then prepared by amplification with 2x KAPA HiFI HotStart mix and NEB dual barcode. Library quality was measured on an Agilent 2100 Bioanalyzer (DNA 1000 chip) for concentration and product size. Paired-end libraries were sequenced with the Illumina NovaSeq, (2 × 151 nucleotide read length) with sequence coverage to 30-40 million paired reads.