GSM5822262: pre-B cells RAG-/- u+ H3K9me3 WT_2; Mus musculus; OTHER
CUT& RUN was from (Hainer and Fazzio, 2019) with some modifications. 33.3 ul of Concanavalin A beads (Bangs Laboratories, cat# BP531) were used per 1x106 cells. 1-3x106 cells were used per sample. Incubation of nuclei-bound beads with antibody was done overnight at 4°C on a rotating platform. All antibodies were used in a 1/100 dilution. DNA purification after fragment release was done using NucleoSpin Gel and PCR Clean‑up kit (Ref#740609.50, Macherey-Nagel) instead of phenol-chloroform-isoamyl solution extraction. CUT&RUN libraries for illumina sequencing were prepared with KAPA HyperPrep Kit (KK8502) using KAPA Unique Dual-Indexed (UDI) Adapters (KK8727).