GSM3693232: Male_Heart_DroNc; Homo sapiens; RNA-Seq
For Drop-seq, 2 mL of cells at 200,000 cells/mL in PBS-BSA was loaded in a 3 mL syringe (BD #309657). A custom-built 90 μm Drop-seq microfluidic device (CAD file supplied separately) was used for droplet generation, creating droplets smaller than the standard Drop-seq protocol. For DroNc-seq, a 75 μm microfluidic device9 was used. 2 mL of nuclei at 300,000 nuclei/mL were loaded into a 3 mL syringe and flowed at 1.5 mL/hr (similar parameters were used for nuclei from primary heart tissue). Drop-seq and DroNc-seq samples for each differentiation time-point were sequenced in a single run, with 150-200 million reads allocated per sample. Sample libraries were loaded at ~1.5 pM concentration and sequenced on an Illumina NextSeq 500 using the NextSeq 75 cycle v3 kits for paired-end sequencing.
GEO Accession
GSM3693232