GSM7856996: ROI1_genomic_DNA_FC_biorep1_techrep1_P3; Homo sapiens; OTHER
gDNA extraction: gDNA was extracted from approximately 3-4 million cells with the Cell and Tissue DNA Isolation Kit (Norgen Biotek Corp, 24700), including RNaseA treatment at 37 ˚C for 15 min, and eluting in 200 µL warm elution buffer.  RNA extraction: Approximately 3-4 million cells were solubilized with 1 mL QIAzol (Qiagen, 79306) and 0.2 mL chloroform in 5PRIME Phase-Lock Gel heavy tubes (QuantaBio, 2302830), according to the manufacturer's recommendations. RNA was precipitated at -20 ˚C following the addition of 2 µL GlycoBlue (Thermo Fisher, AM9515) and 2.5 volumes of cold absolute ethanol. RNA was washed once with cold 70% ethanol then resuspended in 30 µL water. 10 µg total RNA or 15 µL of polysomal RNA (half of 30 µL eluate, volumetric inputs) was treated with DNaseI (NEB, M0303) in a 100 µL reaction at 37 ˚C for 15 min, then re-purified by the RNA Clean and Concentrator Kit (Zymo Research, R1015) and eluted in 20 µL water.  Polysome fractionation and polysomal RNA purification: For COMT library stable cell lines, cells at 80-95% confluency (one 15 cm dish per gradient) were incubated with 100 µ