GSM7035130: H3K27me3_rep2; Mus musculus; ChIP-Seq
Olfactory tissue was obtained from adult wild-type mice euthanized 7-10 days following methimazole lesion by decapitation after deep anesthesia, and olfactory mucosa was dissociated in an enzyme cocktail (collagenase, Dispase and DNase I in HBSS for 15 min, followed by 3 min with 0.125% trypsin with gentle trituration. Cells from 3 mice were pooled per ChIP replicate, and GBCs were purified from the cell suspension using APC-conjugated antibody to c-Kit for 15 minutes and then APC magnetic selection. Cells were crosslinked in 1% formaldehyde for 10 minutes at room temperature.Cells were lylsed and homogenized, and a Bioruptor PIco was used to shear chromatin. Chromatin was incubated with antibodies (30 μg chromatin and 4 μg of antibody) at 4°C overnight. NEBNext Ultra II DNA Library Prep Kit for Illumina with at least 1 ng of ChIP genomic DNA was used as starting material for library prep following manufacturer instructions.