GSM5289681: iSABs; Mus musculus; RNA-Seq
iSABs were separated using the Primary Cardiomyocyte Isolation Kit (Thermo Fisher Scientific) and resuspended in PBS with 0.04% BSA to 1.000 cells/μl. Single cells were then captured in droplet emulsions using the GemCode Single-Cell Instrument (10x Genomics) with a target output of 2.000 cells. Libraries for scRNA-seq were constructed according to the 10x Genomics protocol using the GemCode Single-Cell 3′ Gel Bead and Library V3 Kit. Amplified cDNA and final libraries were evaluated on the 2100 Bioanalyzer instrument (Agilent) using a High Sensitivity NGS Analysis Kit (Advanced Analytical). Amplification yielded in 3.3pg/µl cDNA and final libraries contained cDNA fragments with an average size around 450bp. The subsequent sequencing was conducted on the HighSeq4000 Sequencing System using the HiSeq SBS and HiSeq PE Cluster Kit V4 (all Illumina, San Die-go, CA. USA).