GSM6797043: CUT&RUN H3K27ac primary RPTEC, rep 2; Homo sapiens; OTHER
CUT&RUN was performed with the CUTANA kit (Epicyper, 14-1048). Cell cultures were fixed with 0.5% formaldehyde for 1 minute at room temperature, then quenched with glycine to a final concentration of 125mM. Cells were scraped from dishes then spun down at 500g for 5 minutes. Cell pellets were resuspended in PBS with 1% BSA, counted, spun down again, and resuspended again. 500,000 cells in 100 ul wash buffer were incubated with Concanavalin A-conjugated paramagnetic beads, then antibodies (1:50) were added to each sample. Remaining steps were manufacturer instructions for cross-linked samples. Library preparation was performed with NEBNext ULTRA II DNA Library Prep Kit for Illumina (New England BioLabs, E7645S)