GSM8432353: 50nM siSptbn1, replicate 2, RNAseq; Homo sapiens; RNA-Seq
snRNA-seq: Nuclei were isolated by lysing frozen mouse livers with a pestle and cold lysing buffer (10mM Tris-Cl, PH 7.4, 10mM NaCl, 3mM MgCl2, 0.1% NP-40, 0.4 U/ul RNase Inhibitor, 1mM DTT) for 4 min on ice. The lysate was filtered over a 40um filter, centrifuged at 500xg at 4 deg and resuspended in 1X PBS with 1% BSA & 0.4 U/ul RNase Inhibitor. snRNA-seq: Nuclei were labeled with 7-AAD (Sigma) and sorted on a BD FACS Aria to purify nuclei from the remaining debris. Sorted nuclei were counted and loaded onto a Chip G and 10X Genomics Chromium Controller to generate GEMs following the manufacturer's protocol (Chromium Next GEM Single Cell 3'v3.1 with Dual Indexing). Libraries were quantitated and fragment sizes determined using a High Sensitivity DNA Chip and Bioanalyzer (Agilent). RNA-seq: RNA was extracted from liver tissues using the RNeasy Plus Mini kit. RNA quality and concentration were assessed using the Thermo Scientific Nanodrop spectrophotometer before RNA sequencing. RAN-seq: About 1 μg of RNA was used to generate barcoded sequencing libraries. Novogene Corporation performed library preparation.