GSM8291977: P100_cont1; Mus musculus; RNA-Seq
Nuclei were isolated with Nuclei EZ Lysis buffer (NUC-101; Sigma-Aldrich) supplemented with protease inhibitor (5892791001; Roche) and RNase inhibitor (Protector RNase Inhibitor). Samples were cut into < 1 mm pieces, homogenized using a Dounce homogenizer (885302–0002; Kimble Chase) in 2 ml of ice-cold lysis buffer, and incubated on ice for 5 min with an additional 1 ml of lysis buffer. The homogenate was filtered through a 40-μm cell strainer and centrifuged at 500 g for 5 min at 4°C. The pellet was resuspended, washed with 3 ml of buffer, and incubated on ice for 5 min. Following centrifugation, the pellet was resuspended in Nuclei Buffer (10× Genomics, PN-2000153), filtered through a 5-μm cell strainer, and counted. All these processes were performed in a cold room at 4°C. Subsequently, nuclei suspension was diluted to target 10,000 nuclei per lane and loaded into a thermal cycler to allow transposition reaction. Chromium Next GEM Single Cell Multiome ATAC+ Gene Expression Reagent Kit (10X Genomics) Single nucleus multiomics with simultaneous snRNA-seq + snATAC-seq with multiome kit [10XGenomics]