GSM8395409: Healthy Subject 5,Nasal Airway cells, scRNAseq; Homo sapiens; RNA-Seq
Airway cells were isolated after incubation in trypsin for 5 minutes and mechanical dissociation. The isolated cells were resuspended in tryspin for  and incubated at 37 °C for 5 min. Cells were washed and re-suspended in PBS and stained with a Viability Dye, washed and resuspended in FACS buffer (PBS+0.04% BSA). Library was performed according to the manufacter's instructions (single cell 3' v2 protocol, 10x Genomics). Briefly, cells 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 a primer sequence, Unique Molecular Identifier (UMI) and the 10x Barcode. The pooled barcoded cDNA was then cleaned up, amplified, and the apropiated sized fragments were selected for subsequent library construction.