GSM8863501: Whole mouse heart after PBS inhalation treatment (control) (hearts from multiple mice); Mus musculus; RNA-Seq
The sample was first prepared as a single cell suspension by using gentleMACS and associated tissue kit, followed by cell counting and cell viability assay. If the cell viability was 80%, the cell concentration was adjusted to 1000/µL. The prepared cell suspensions were captured as a cell and a bead with a cell Barcode in a droplet using a microfluidic chip, and a droplet containing the cell was collected. In the droplet, the cells were lysed so that the mRNA in the cells was connected to the cell Barcode on the bead to form Single Cell GEMs (Gel Bead-in-Emulsions), and an RT reaction was carried out in the droplets, followed by demulsification to construct a cDNA library. The cell Barcode on the library sequence distinguished which cell the target sequences were from, and the sample Index on the sequence distinguished which sample the target sequences were from.  The raw data obtained by sequencing is called raw reads. We split the Barcode, UMI and insert parts of reads according to the unique library structure of 10x scRNA-Seq, and then inserted the fragments for comparison to the reference genome. Then, the ratios of the comparisons to the respective regions were statistically analyzed, and the expression amount statistics were performed; based on the results of the expression amount, the cell cluster analysis and the screening and identification of the marker genes were performed.