GSM7232734: 1545-H3K4me3; Homo sapiens; OTHER
100,000 cells per line per antibody / condition was washed with 1X Wash Buffer three times, before resuspension in 100 µl of 1X Wash Buffer (+ spermidine + PIC), according to protocol. CUT&RUN (Cleavage Under Targets & Release Using Nuclease) libraries were prepared with the established CUT&RUN Assay Kit (Cell Signaling, #86652S). CUT&RUN libraries were prepared using the SimpleChIP ChIP-seq DNA Library Prep Kit for Illumina (Cell Signaling, #56795S) with SimpleChIP ChIP-seq Multiplex Oligos for Illumina (Cell Signaling, #47538S) according to manufacturer guidelines, with modifications for CUT&RUN as noted in the CUT&RUN Assay Kit (Cell Signaling, #86652S). All libraries were quantified on the Qubit 4.0 and library quality assessed using Agilent High Sensitivity DNA Tapestation (Agilent, 5067-5584) before multiplexing and sequencing on a Nextseq 550 Mid Output Kit (150 cycles) with 1% PhiX for a minimum of 5 million paired-end reads per sample with 50bp read length.