GSM6505392: control 1; Rattus norvegicus; RNA-Seq
Nuclei were extracted from frozen lung tissue using Singulator TM 100 (S2 Genomics), with the following settings: nuclear reagent, S2; incubation temp, cold; mixing type, top; mixing speed,  fastest; disruption type, default; disruption speed, medium. RNase inhibitor was added to the Singulator Cartridge to reach a final concentration of 0.2U/μl. Nuclei collected from the Singulator was spun at 800g, at 4°C for 10min. Supernatant was removed, following by resuspending nuclei in freshly made cold Wash and Resuspension Buffer (1X PBS, 1% BSA, 0.2U/μl). The wash was repeated twice before proceeding into the 10X Genomics single-cell 3' v3.1 assay and processed as described by the protocol provided by the 10X Genomics. Libraries construction and library sequencing were proceeded as described in the 10X Genomics protocol using NovaSeq 6000.