GSM6310595: Epithelial_K5_scATAC1; Mus musculus; ATAC-seq
FACS-sorted cells were lysed in cold-lysis buffer (0.1% NP-40, 0.1% Tween 20, 0.01% Digitonin, 10 mM NaCl, 3 mM MgCl2 and 10 mM Tris-HCl [pH 7.4]), washed and processed according to “Nuclei Isolation for Single-Cell ATAC Sequencing” protocol (CG000169 RevD). Resulting nuclei suspension was subjected to transposition reaction for 60 min at 37°C and then encapsulated in microfluidic droplets using 10X Chromium instrument following manufacturer's instructions with a targeted nuclei recovery ~ 5,000. Barcoded DNA material was cleaned and prepared for sequencing according to the Chromium Single Cell ATAC Reagent Kits User Guide (10x Genomics; CG000168 RevA). Purified libraries were assessed using a Bioanalyzer High-Sensitivity DNA Analysis kit (Agilent) and sequenced on a Illumina HiSeq 2500 (High Output) platform at approx. 150M reads (R1 50bp, R2 50 bp, i7 8bp, i5 16bp) per 1 sample (~ 5000 nuclei) at MSKCC's Integrated Genomics Operation Core. 