title: GSM2065674: Wild-type Lgr5+ ISC H3K27me3 ChIP-Seq rep 2; Mus musculus; ChIP-Seq
library_construction_protocol: 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 ThruPLEX DNA-seq Kit (Rubicon Genomics) was used to prepare libraries, and 75 bp single-end reads were sequenced on an Illumina NextSeq 500 instrument.
attributes.tag: GEO Accession
attributes.value: GSM2065674