GSM1843533: Wild-type Lgr5+ ISC Input DNA; 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, but not immunoprecipitated. 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.