GSM9788323: CBO, EB120, Rep 2; Homo sapiens; RNA-Seq
CBOs at EB60 and EB120 were dissociated using papain/DNase I at 37°C for 30 min and triturated into single-cell suspensions. Cells were pelleted and lysed for 3 min in cold lysis buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20, 0.1% Igepal CA-630, 0.01% digitonin, 1% BSA), then washed with chilled wash buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20, 1% BSA), and nuclei were pelleted at 500 rcf for 5 min at 4°C Single-cell libraries were constructed using the Chromium Next GEM Single Cell 3ʹ Kit v3.1 (10X Genomics), targeting 10,000 cells per sample. GEM-based barcoding was followed by reverse transcription to generate 10X barcoded cDNA.