title: GSM6928478: scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: Cardiac Lin- CDCs were isolated from the hearts of 2- to 3-month-old male C57BL/6 mice. Briefly, in step 1, mouse hearts were perfused with an enzyme solution using the Langendorf Perfusion Isolation System for Adult Mouse Cardiomyocyte Isolation Kit according to the manufacturer's protocol. In step 2, digested cells are filtered through a 40 μM cell strainer to remove the majority of cardiomyocytes and then sorted by magnetically activated cell sorting (MACS) using the Mouse Lineage Cell Depletion Kit. Selected Lin- CDCs were sent to Genome Core for scRNA-seq library preparation. Library was performed according to the manufacter's instructions (10x Genomics). Briefly, CDCs were resuspended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in-emulsion (GEM). The poly-A RNA from the cell lysate contained in every single GEM was retrotranscripted to cDNA, which contains an Ilumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10x Barcode. The pooled barcoded cDNA was then cleaned up with DynaBeads, amplified by PCR and the apropiated sized fragments were selected with SPRIselect reagent for subsequent library construction. During the library construction Ilumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added.