GSM6556450: Human Control Liver 2, snRNA-seq; Homo sapiens; RNA-Seq
40-60mg of total liver tissue  was chopped on ice in 1mL of 0.03% TST buffer (146mM NaCl, 10mM Tris-HCl pH 7.5, 1mM CaCl2, 21mM MgCl2 with freshly added 0.03% Tween-20, 0.01%BSA, and 0.4U/mL Ribolock RNase Inhibitor (Thermo Fisher Scientific FEREO0382, MA, USA)) [30] with Tungsten Noyes Spring Scissors (FST 15514-12, CA, USA). The resulting nuclei suspension was then filtered through 40mm cell strainers (Thermo Fisher Scientific 352340, MA, USA) into a fresh 50mL conical tube on ice. 1 more mL of 0.03% TST was used to rinse the cell strainer and 3mL of ST buffer was added to the nuclei suspension followed by briefly mixing by gentle flicking. The nuclei suspensions were spun at 500xg for 5 minutes at 4°C in a swing-bucket centrifuge. The final nuclei pellet is suspended in 200mL of 0.4% BSA in PBS by pipetting 30 times with a regular p1000 tip. Nuclei preparations were processed by the Chromium 3′ Gene Expression V3 Kit according to the manufacturer's guidelines. Qubit 3 (Fisher Scientific) and 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA) were used for quality check of cDNA.