GSM8455182: Ln, total, GEX; Mus musculus; RNA-Seq
On day 17, lungs were removed and bronchial lymph nodes were dissected from the lung tissue under a stereo microscope. Single-cell suspensions from lung tissue were prepared using the Miltenyi gentleMACS Dissociator and digested with 0.5 mg/ml collagenase D and 20 mg/ml DNase I for 25 min at 37°C. Antigen-specific B cells were first enriched for CD45.1 expressing cells by magnetic sorting. For higher purity, cells were sorted on a Becton Dickinson ARIA II or ARIA Fusion cell sorter as DAPI- CD4- CD8- CD19+ CD45.1+ CD45.2- total B cells or additionally as CD138- CD38low GL7+ geminal center (GC) B cells. Library construction was performed according to 10x Genomics instructions. For samples 1 and 2, the Chromium Next GEM Single Cell 3' v2 Kit was used. For samples 3-6, the Chromium Next GEM Single Cell 5' v2 kit in combination with the Single Cell Mouse BCR Amplification Kit was used.