GSM8655794: Myc-Cap, DMSO treated - 1; Mus musculus; RNA-Seq
Total RNA was extracted using the RNeasy Mini Kit (Qiagen) and quantified by NanoDrop 2000 Spectrophotometers (Thermo Scientific). RNA-seq libraries were prepared using KAPA RNA Hyper+RiboErase HMR (Roche Cat#: 08098140702). Ribosomal RNA was removed by enzymatic digestion from 800 ng total RNA, following with cDNA and double strand cDNA synthesis, end repair, A-tailing, and ligation with NEB adapters. cDNA fragments with sizes between 250-300 bp were separated using double AMPure beads and PCR amplified using 2x KAPA HiFi HotStart mix and NEB dual indexes (Roche Cat#: E6440L). Library quality was measured on an Agilent 2100 Bioanalyzer for concentration and product size. Paired-end libraries were sequenced with the Illumina NovaSeq 6000 (2 × 150 nucleotide read length) with sequence coverage to 20-30 million paired reads.