title: GSM9220032: cardiac organoid at day 36; Homo sapiens; ssRNA-seq
library_construction_protocol: Cardiac organoid samples were dissociated via a cold activated B. Licheniformis protease Library preparation was performed according to the manufacturer's instructions (Chromium Next GEM Single Cell 3' v3.1 protocol CG000390 Rev B, 10x Genomics). Briefly, cells were resuspended in the master mix and loaded, along with partitioning oil and gel beads, into the microfluidic chip to generate a gel beademulsion (GEM). The poly-A RNA from the cell lysate contained within each droplet was reverse transcribed into cDNA, which included an Illumina R1 primer sequence, a Unique Molecular Identifier (UMI), and the 10x Barcode. The pooled, barcoded cDNA was then purified using Silane DynaBeads, amplified by PCR, and size-selected using SPRIselect reagent to isolate appropriately sized fragments for subsequent library construction. During library construction, the Illumina R2 primer sequence, paired-end constructs with P5 and P7 sequences, and a sample index were added. Library quantification and quality control were performed using a Qubit™ 4.0 Fluorometer (Thermo Fisher) and a 2100 Bioanalyzer with the High Sensitivity DNA Kit (Agilent). Sequencing was carried out on a NovaSeq 6000 sequencer (Illumina) using an S2 flow cell.