GSM6190727
To prepare nuclei, around 50,000 cells at 500g for 5 min, which was followed by a wash using 50 μL of cold 1× PBS and centrifugation at 500g for 5 min. Cells were lysed using cold lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2 and 0.1% IGEPAL CA-630). Immediately after lysis, nuclei were spun at 500g for 10 min using a refrigerated centrifuge.  For mouse: Briefly, cells were fixed with 0.1% formaldehyde and incubated at room temperature for 5 min. The fixation was stopped by adding glycine to the final concentration of 125 mM. The sample was incubated at room temperature for 5 min and washed in PBS. The cell concentration was counted, and approximately 1600-2000 cells per well were distributed into each well of a 96-well plate. Cells were transposed with 96 uniquely barcoded Tn5 at 37°C for 30 min with shaking at 300 rpm. The reaction was stopped by adding 0.5 M EDTA and incubated at 37°C for 15 min. All the cells were then pooled and MgCl2 was added to the pooled sample to quench EDTA. The sample was re-distributed onto another 96-well plate with 20 cells in each well by FACS sorting. Reverse crosslinking buffer and barcode PCR primers were added to each sample. The plate was incubated at 55°C for 16 hours for reverse crosslinking. Tween 20 was then added to quench SDS before PCR amplification. The PCR reaction was carried out at the following conditions: 72°C for 5 min (extension), 98°C for 5 min, and then thermocycling at 98°C for 10 s, 70°C for 30 s and 72°C for 1 min for 12-15 cycles. Libraries were pooled and purified using Qiagen MinElute PCR purification column. The libraries were quantified using KAPA library quantification kit. Libraries were sequenced on the Next-seq platform (Illumina) using a 150-cycle kit (Read 1: 47 cycles, Index 1: 36 cycles, Index 2: 36 cycles, Read 2: 47 cycles). For marmoset: Somatosensory and motor cortex from wild-type animals was dissected and dissociated with the Worthington Papain Dissociation System (Worthington Biochemical Corporation), and live cells were isolated by FACS sorting as DAPI-negative, Vybrant DyeCycle Ruby (Thermo Fisher)-positive events. Libraries were prepared using the 10x Genomics Chromium Single Cell ATAC kit (10x Genomics).