GSM8409469: tdTomato- cDCs from naïve Il7r-cre; rosa26-lsl-tdTomato mice; Mus musculus; RNA-Seq
Spleens were harvested from naïve Il7r-cre; Rosa26-lsl-tdTomato mice and cDCs were enriched by using magnetic cell sorting with APC-labeled anti-CD11c antibody and anti-APC microbeads. Then, cells were stained and L-cDCs and M-cDCs were isolated by FACS sorting. Cells were washed and re-suspended in PBS. Single cell gene expression analysis was conducted by GENEWIZ from Azenta Life Sciences (Tokyo, Japan). Approximately 10,000 cells with a 90% survival rate were applied to GEM (Gel Bead-in-Emulsion) generation using Chromium Controller (10x Genomics, Pleasanton, CA, USA), where individual cells were captured into single oil emulsions with reverse transcription reagent and a Gel Bead containing barcoded oligonucleotides. The reverse transcription reaction enables individual cDNAs to be labeled by unique cell barcodes with unique molecular tags in each single cell emulsion. cDNA amplification followed by library construction for single cell gene expression was performed using a Chromium Next GEM Single Cell 3ʹ Kit v3.1 (10x Genomics). The resulting double-stranded libraries were quantified by Qubit dsDNA Assay (Thermo Fisher Scientific) and TapeStation D1000 ScreenTape (Agilent Technologies), then converted and processed into the form of single-stranded circular DNA (sscDNA) in order to be analyzed on a MGI sequencing platform.