GSM4970301: H28126 - 24 weekATAC-seq; Homo sapiens; ATAC-seq
Cerebral organoids were dissociated in Papain containing Deoxyribonuclease I (DNase) for 30 minutes at 37C and samples were triturated to form a single cell suspension. For scATAC-seq, cells were pelleted and lysed for 3 minutes in 100uL chilled Lysis Buffer (10mM Tris-HCl pH7.4, 10mM NaCl, 3mM MgCl2, 0.1% Tween-20, 0.1% Igepal CA-630, 0.01% Digitonin, 1% BSA). Lysed cells were then washed with 1mL chilled Wash Buffer (10mM Tris-HCl pH7.4, 10mM NaCl, 3mM MgCl2, 0.1% Tween-20, 1% BSA) and nuclei were pelleted at 500rcf for 5 minutes at 4C. Nuclei were prepared as outlined in the 10X Genomics Chromium single cell ATAC-seq solution protocol (v1.0 kit was used). Nuclei were loaded with a capture target of 10,000 nuclei per sample. scATAC-seq libraries were prepared for sequencing following the 10X Genomics single cell ATAC-seq solution protocol. scATAC-seq libraries were sequenced using PE150 sequencing on an Illumina NovaSeq with a target depth of 25,000 reads per nucleus.