GSM7789731: IgG rep1; Mus musculus; OTHER
Mammary gland tissue was dissociated using gentle collagenase/hyaluronidase (Stemcell), mammary epithelial cells were further isolated using 0,25 % trypsin (Gibco) and 5 mg/mL dispase  (Merck). Mammary epithelial luminal cells were sorted via FACS using EpCAM and CD49f surface markers (antibodies:  12-9326-42 from eBioscience and 313631 from Biolegend). For each CUT and RUN reaction, 200,000 cells were bound to activated concanavin A magnetic beads (Polysciences) and incubated with the desired antibody (1:100) in the presence of 0.01 % digitonine. DNA was cleaved using 1 μg/mL protein A/G Micrococcal Nuclease fusion protein (pA/G MNase) in the presence of 0.01% digitonine. Samples were treated with RNAse and Protreinase K and the released DNA was purified using Phenol/Chloroform/Isoamyl alcohol DNA library generation with the NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® (New England Biolabs) according to the manufacturer's recommendations. Adaptor ligation was performed with 1:25 diluted adaptor and 15 cycles were used for library amplification.