title: GSM7868273: LM_snRNA_seq; Mus musculus; RNA-Seq
library_construction_protocol: FFPE samples were preprocessed on a prototype Singulator™ system. The sample was automatically processed in a NIC+™ cartridge (S2 Genomics #100-215-389) by three 10 min deparaffinization steps (CitriSolv, VWR), rehydrated by successive 1 mL washes of 100%, 100%, 70%, 50%, and 30% ethanol, followed by 2 washes of PBS. The sample was then spun at 1,000g for 3 min and resuspended in 0.5 mL Nuclei Isolation Reagent (NIR, S2 Genomics, #100-063-396) with 0.1 u/mL RNase inhibitor (Protector™, Millipore Sigma, #3335399001); all subsequent solutions had RNase inhibitor. The sample was dissociated to single nuclei in a second NIC+ cartridge with 2 mL of NIR for 10 min followed by a 2 mL wash with Nuclei Storage Reagent (NSR, S2 Genomics, #100-063-405). The single nuclei suspension was spun 500g for 5 min, resuspended in NSR, and counted. Single-nuclei RNA-Seq of FFPE-isolated nuclei suspensions was performed on Chromium instrument (10X genomics) following the user guide manual for Chromium Fixed RNA Kit, Mouse Transcriptome (SinglePlex) Illumina NovaSeq S4 platform (R1 – 28 cycles, i7 – 8 cycles, R2 – 90 cycles)