GSM4928831: 4CRE; Mus musculus; ATAC-seq
Subcutaneous tissue was isolated and adipose deposits was removed under sterile conditions and washed in PBS supplemented with 100 U/mL penicillin and 100 μg/mL streptomycin on ice. Tissue was minced and digested with 1 mg/ml collagenase type I and 0.5%Trypsin in 0.1% BSA, for 70 minutes at 37 °C. Digested tissue was centrifuged at 500 g for 10 min and the pellet was carefully collected after aspirating off the floating fat depots. After a second centrifugation at 500 g for 10 min, the cellular pellet was filtered through a 100-μm mesh filter to remove debris. Nuclei were isolated from 50,000 cells for transposition reaction with Tn5 Trnsposase (Illumina, catalog # FC-121-1030). DNA were purified by using PCR purification kit (Qiagen). To generate sequencing libraries, purified DNA was amplified with NEBNext high-fidelity 2X PCR Master Mix (New England Biolabs). Sequencing libraries was sequenced using Illumina Nextseq 550 sequencer to obtain 150bp paired-end reads.
GEO Accession
GSM4928831