GSM9037912: H2A.Z CUT&Tag in ZX-1, Dox, rep5; Mus musculus; OTHER
Cells were bound on ConA beads and incubate with the primary antibody or proteins overnight in antibody buffer. For CUT&Tag, cells were incubated with secondary antibody and pA-Tn5 sequentially and then tagmented with tagmentation buffer. Tagmentation was stopped by EDTA buffer and DNA were then recovered and directly subjected to library amplification. For CUT&RUN, cells were incubated with pA-MNase and digested on ice-water in buffer containing CaCl2. Digestion was stopped by 2X STOP buffer and DNA were recovered by high speed spin, Proteinase K/SDS digestion and PCI extraction. For MapR, cell were directly applied for digestion and the rest step as described in CUT&RUN. For CUT&Tag, tagmented DNA was recovered and directly PCR with Nextera primer for library amplification. For CUT&RUN and MapR, NEBNext Ultra II DNA library prep was used for library construction with 14 cycle of library amplification.