title: GSM9561418: Human Brain Metastasis Tissue, LBM-RIH-3, Multiome, GEX; Homo sapiens; RNA-Seq
library_construction_protocol: Fresh Brain Metastasis tissue samples were collected, transferred to a tube containing MACS© tissue storage solution (Miltenyi) and placed on ice before being processed for tissue snap freezing, OCT cryopreservation or cell dissociation within 24h. The proportion of each tissue sample designated for scMultiome-seq was snap frozen in liquid nitrogen and stored at -80C until nuclei isolation. Nuclei isolation from frozen BM patient tissue samples was performed according to the 10x Genomics 'Nuclei Isolation from Complex Tissues for Single Cell Multiome ATAC + Gene Expression Sequencing' protocol. Nuclei isolation from CAF cell lines was performed according to the 10x Genomics 'Nuclei Isolation for Single Cell Multiome ATAC + Gene Expression Sequencing'. Nuclei quality check and concentration measurement was performed at the end of the isolation process using a Countess II FL Automated Cell Counter (ThermoFisher Scientific) and ApoTome.2 microscope (ZEISS). Nuclei suspensions were loaded onto a 10x Genomics Chromium Next gel beads in emulsion (GEM) Chip for transposed nuclei capture using 10x Genomics Chromium Controller. ScMultiome-seq libraries were performed using Chromium Next GEM Single Cell Multiome Reagent Kits according to the 'Chromium Next GEM Single Cell Multiome ATAC + Gene Expression' user guide provided by 10x Genomics.