GSM8403666: T8_ARC_150; Mus musculus; RNA-Seq
Left atrial tissues from vehicle- and anti-Trem2-siSpp1-treated HOMER mice were microdissected using a dissection microscope and subsequently snap-frozen in liquid nitrogen. Nuclei were then isolated using the 10X Chromium Nuclei Isolation kit according to the manufacturer's protocol. In detail, frozen samples were homogenized using a pestle and lysis buffer and passed through a column. Next, debris was removed via centrifugation in debris removal buffer. The nuclei were then washed, resuspended and visualized under a microscope to determine nuclei concentration and quality. The isolated nuclei were immediately loaded on a 10X Genomics Chromium Single Cell G chip as outlined by the 10X Genomics Chromium Single Cell 3' Reagent kit v3.1 user guide. As nuclei have a lower RNA content than whole cells, the cDNA amplification cycles were increased by 1-2 cycles, as recommended by 10X Genomics. Libraries were prepared as outlined by the 10X Genomics Chromium Single Cell 3' Reagent kit v3.1 user guide with appropriate modifications to the PCR cycles based on the cDNA concentration. Constructed libraries were validated and quantified with the Agilent High Sensitivity DNA chip and by real-time qPCR and subsequently sequenced on an Illumina NovaSeq 6000 system as recommended by 10X Genomics. snRNA-seq