GSM9278511: Multiome_Control_Heart_3months; Mus musculus; RNA-Seq
Snap-frozen mouse hearts were thawed and prepared for nuclei isolation. Tissues were washed in 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; 1 U/μl RNase inhibitor), centrifuged, 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; 1 U/μl RNase inhibitor). Samples were lysed on ice for 5 min and resuspended in 1 ml chilled wash buffer. Following centrifugation at 500 rcf for 5 min, the nuclei pellet was resuspended in 50 μl 10x Genomics Nuclei Buffer containing 1 U/μl RNase inhibitor. For the Gene Expression arm, polyadenylated nuclear RNA was reverse-transcribed within GEMs to produce barcoded cDNA (cell barcode and UMI), followed by cDNA amplification and 3′ gene-expression library construction (fragmentation, end-repair/A-tailing, adapter ligation, and sample index PCR). For the ATAC arm, transposition was performed with Tn5 to fragment accessible chromatin and attach partial adapters; after GEM recovery, barcoded ATAC fragments were amplified and indexed.