GSM8900545: Multiome, murine lung tissue, GEX4; Mus musculus; RNA-Seq
Nuclei were isolated from human snap frozen lung tissue using the 10x genomics nuclei isolation kit, per manual (document number CG000505, revision A, May 2022). An aliquot of nuclei was stained with 0.4% trypan blue stain and DAPI and subsequently counted with the Thermo Fisher countess II FL. Samples with > 90% viability were used for downstream analysis. Approximately 10,000 nuclei per sample were loaded onto chip J and processed with the Chromium X Series to create to create Gel Beads-in-emulsion (GEMs). The next steps were carried out according to the manufacturer's user guide (CG000338, revision G, September 2024) [10], including post-GEM-RT Dynabead cleanup and Pre-Amplification. Each sample was then split into two portions: one for ATAC library construction and the other for cDNA amplification and gene expression library construction. DNA from the ATAC libraries (scATAC libraries) and cDNA from the gene expression libraries (scRNA libraries) were assessed at two time points using the Agilent Bioanalyzer High Sensitivity Chip to ensure quality. Six scATAC libraries and six scRNA libraries were sequenced on the Novaseq 6000 platform, using 2 × 100 bp paired-end sequencing configuration.