GSM1843532: Wild-type Lgr5+ ISC H3K27me3 ChIP-Seq rep 1; Mus musculus; ChIP-Seq
Crypts from the proximal 1/3 of mouse small intestines were isolated by scraping off villi and incubating in 5mM EDTA in PBS for 45 minutes at 4°C. Crypts were disaggregated with 4X TrypLE in DMEM for 45 minutes at 37°C and Lgr5-GFP-HI intestinal stem cells were sorted by flow cytometry. Cells were snap-frozen in liquid nitrogen and pooled for subsequent ChIP analysis. Whole cell lysates were sonicated and incubated with 5 ug H3K27me3 antibody overnight (07-449, Millipore) for immunoprecipitation. The ChIPseq DNA Sample Prep Kit (Illumina) was used to prepare libraries, and 50 bp single-end reads were sequenced on an Illumina Hi-Seq 2000 instrument.