title: GSM6460304: Pdgfra_lo_CD81-rep2; Mus musculus; ATAC-seq
library_construction_protocol: Adult murine small intestinal tissue was harvested, washed and the outer muscle layer was removed with tweezers under a stereomicroscope. Tissue pieces were incubated twice in 10mM EDTA in HBSS  +10mM + 5%FBS for 30min under 300rpm shaking at 37°C, washed and digested with 3mg/ml collagenase2 for 20min at 37°C. The filtered cell suspension was stained with biotinylated anti-CD81 (eBiosciences 13-0811-81, 1:50) or control biotinylated IgG for 25min at 4°C, washed and incubated with streptavidin-APC (eBiosciences, 17431782, 1:100) for 25min at 4°C. Cells were sorted three-way for Pdgfra_eGFP high; Pdgfra_eGFP low CD81+ and Pdgfra_eGFP low CD81-  at the Dana-Farber Flow cytometry core facility using a BD FACS M Aria II instrument. ATAC-seq was performed on 50,000 freshly sorted live mesenchymal cells using the “Omni-ATAC” protocol as described in Buenrostro, J. D., Wu, B., Chang, H. Y., & Greenleaf, W. J. (2015). Protocols in Molecular Biology, 109, 21.29.1-9. https://doi.org/10.1002/0471142727.mb2129s109 and library preparation as described in. Library quantification was performed by Qubit dsDNA high-sensitivity assay and library quality was assessed on a High Sensitivity DNA Kit on a Bioanalyzer 2100. Libraries were size-selected to exclude primer via AMPure beads