GSM8830669: LSK, HFD, H3K4me3, rep2; Mus musculus; OTHER
The bone marrow cells were isolated and separated by magnetic sorting and subsequently further sorted by flow cytometory sorting. Cells were collected in 30% fetal bovine serum containing RPMI media. After washing, nucleus is isolated followed by CUT&Tag using a custom kit. We performed CUT&Tag using a modified protocol based on Kaya-Okur et al. (2019) and consults with Dr. Henikoff (Fred Hutch). Approximately 10,000 cells were incubated with primary antibodies, followed by secondary antibodies and pA-Tn5 fusion protein. After tagmentation, DNA was extracted and amplified by PCR for 14 cycles. Libraries were sequenced on an Illumina NextSeq 2000 platform with paired-end 75 bp reads.