GSM5970641: IgG_Ctrl; Mus musculus; OTHER
Live c-kit+ cells from chimeric mice were stained using Zombie Violet fixable viability kit (Biolegend) and APC-c-kit (Biolegend clone 2B8). Sorting for live c-kit+ cells was performed using a FACSAria III cell sorter (BD Biosciences). CUT&RUN was performed on equal numbers of c-kit+ cells from each group of chimeric mice using the CUT&RUN Assay Kit (Cell Signaling, Kit #86652) per manufacturer's protocol. Chromatin associated with H3K4me3 (Cell Signaling clone C42D8), and rabbit IgG negative isotype control (Cell Signaling clone DA1E) were isolated. Technical replicates were performed in duplicate. CUT&RUN libraries were prepared using Swift S2 Acel reagents on a Beckman Coulter Biomek i7 liquid handling platform from approximately 1ng of DNA according to manufacturer's protocol and 14 cycles of PCR amplification.  Subsequently, libraries were sequenced on a NovaSeq with 150bp paired-end reads.