title: GSM7093997: HCC_T_H30; Homo sapiens; ATAC-seq
library_construction_protocol: Nuclei isolation was performed following the 10x Genomics protocol (Nuclei isolation for single cell ATAC sequencing, revision D). Cells were briefly thawed in a 37 °C water bath and resuspended in 10ml RPMI media supplemented with 10% FBS. After centrifugation at 300 rcf for 5 minutes, the supernatant was removed, and the cell pellet was resuspended in 1ml PBS supplemented with 0.04% BSA and passed through a 40 mm cell strainer. Sample concentration and viability were assessed using the LunaFL fluorescent cell counter. Cells were again centrifuged at 300 rcf for 5 minutes at 4 °C and resuspended in 50ml PBS + 0.04% BSA. After an additional spin at 300 rcf for 5 minutes at 4 °C, 45ml of the supernatant was removed and 45ml of chilled lysis buffer was added to the cell pellet. Cells were incubated in lysis buffer for 20 minutes on ice, after which 50ml of chilled wash buffer was added, and cells were centrifuged at 500 rcf for 5 minutes at 4 °C. 95ml of the supernatant was removed, and nuclei were resuspended in 45ml of chilled 10x Genomics diluted nuclei buffer. After a final centrifugation at 500 rcf for 5 minutes at 4 °C the supernatant was removed, and nuclei were resuspended in a final volume of 7ml of chilled 10x Genomics diluted nuclei buffer. Nuclei were loaded according to the 10x ATAC User guide with a single capture lane per sample. Transposition and nuclei partitioning were completed successfully with uniform emulsion consistency and the GEM incubation PCR was run overnight. All subsequent steps of library preparation and quality control were performed as described in the 10X Genomics ATAC User Guide.