GSM8488148: DRIP_shSETX_66; Homo sapiens; OTHER
Cells were resuspended in 1X TE buffer (Tris HCl pH 7.6 10 mM, EDTA pH 8 1 mM) and lysed O/N at 37°C with 0.5% SDS and 62.5 μg/mlProteinase K in 1X TE buffer. Chromatin was then precipited with isopropanolo and sodio acetato, washed with ethanol and resuspeded in 1X TE for 4h at 30°C. O/N digestion was performed with restriction enzymes cocktail. (Sanz, L. A. & Chédin, F. High-resolution, strand-specific R-loop mapping via S9.6-based DNA–RNA immunoprecipitation and high-throughput sequencing. Nature Protocols 14, 1734-1755, doi:10.1038/s41596-019-0159-1 (2019)) According to Sanz, L. A. & Chédin, F. High-resolution, strand-specific R-loop mapping via S9.6-based DNA–RNA immunoprecipitation and high-throughput sequencing. Nature Protocols 14, 1734-1755, doi:10.1038/s41596-019-0159-1 (2019) Libraries were prepared for Illumina sequencing using a standard protocol consisting in blunting, addition of dA overhangs, ligation of Illumina adapters, selection on gel and PCR with index primers