title: scRNAseq_musmusculus_thymus_mouse_WT_female_4week
design: In vivo Aire WT:In this project, we performed a fresh thymus enzymatic dissociation (treatment with collagenase plus dispase) from an Aire wild-type mouse (C57BL/6 backgound). Thymic stromal cells together with thymocytes and other cell types present in the thymus gland, showed >90% viability. Cells were processed following the 10X Genomics protocol to prepare a single-cell library using Chromium equipment (10X Genomics). The single cells were isolated and lysed. Next, mRNAs were purified and primed with a poly(T) primer for reverse transcription. Unreactive primers were removed by exonuclease I digestion. Poly(A) tails were added to the first strand cDNA at the 3' end and annealed to poly(T) primers for second-strand cDNA generation. The cDNAs were PCR-amplified, sheared, and prepared into sequencing library. The library was sequenced in an Illumina NovaSeq 6000 sequencer.