title: GSM6720576: GEX_sample28; Homo sapiens; OTHER
library_construction_protocol: Fresh cardiac left ventricular tissue from ex-planted hearts at the time of transplantation, LVAD cores, or donors declared DCD were harvested and perfused. Tissues were minced and digested. After incubating with the TotalSeq A 277 panel (BioLegend) antibodies with custom oligo-labeled FAP and LRRC15 antibodies, single cells were isolated and counted before proceeding with the 10x protocol. Collected cells were processed using the single Cell 3' Kit v 3.1 (10x Genomics PN: 1000268). Single-cell libraries were prepared using the single Cell 3' Kit v 3.1 following a modified 3' v3.1 assay protocol (User Guide CG000206) to concurrently prepare gene expression and TotalSeq A antibody derived tag (ADT) libraries as recommended by BioLegend. 1 ul of 0.2uM ADT Additive Primer (CCTTGGCACCCGAGAATT*C*C) and 15 ul of cDNA Primers (PN: 2000089) were used to amplify cDNA. ADT libraries were amplified with a final concentration of 0.25 uM SI Primer (AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGC*T*C) and 0.25 uM TrueSeq Small RNA RPI primer (CAAGCAGAAGACGGCATACGAGAT[6nt index]GTGACTGGAGTTCCTTGGCACCCGAGAATTC*C*A) using 15 cycles. Gene expression libraries were indexed using Single Index Kit T Set A (PN: 2000240). Libraries were sequenced on a NovaSeq 6000 S4 flow cell (Illumina).