title: GSM7094289: Cardiac_Organoid_COVID; Homo sapiens; OTHER
library_construction_protocol: Human iPSC-derived COs were digested with freshly prepared Liberase solution (R&D) for 10 mins. Digestion was halted by resuspending in RPMI-B27 with insulin and EGM2 medium. After passing through a 70 μm strainer, viability of the cells was checked and then prepped for nuclei isolation. Cells were washed with ice-cold Wash buffer (10 mM Tris pH 7.4, 10 mM NaCl, 3 mM MgCl2, 1% BSA, 1 mM DTT, 0.1% Tween-20, and 1 U/μl RNase inhibitor), spun down and resuspended in 100 μl chilled Lysis Buffer (10 mM Tris pH 7.4, 10 mM NaCl, 3 mM MgCl2, 1% BSA, 1 mM DTT, 0.1% Tween-20, 0.01% Digitonin, 0.1% Nonidet P40 Substitute and 1 U/μl RNase inhibitor). Cells were lysed for 5 mins on ice followed by resuspension in 1 ml chilled wash buffer. After spinning for 5 mins at 500 rcf, nuclei pellet was resuspended in 50 μl Nuclei buffer (10x Genomics) and 1 U/μl RNase inhibitor. Briefly, an estimated 10,000 nuclei per sample were captured as per instructions from the10X chromium Single Cell Multiome ATAC and Gene Expression Reagent Kits V1 User Guide. Joint scRNA- and scATAC-seq libraries were prepared using the 10x Genomics Single Cell Multiome ATAC + Gene Expression kit according to manufacturer's instructions