GSM9378460: Sample1 Sample Tag; Mus musculus; OTHER
Splenic single-cell suspensions were generated by mechanical dissociation into supplemented Opti-MEM. Negative enrichment of CD45.1 HkiL B cells was performed using the EasySep Mouse Streptavidin RapidSpheres Isolation Kit (STEMCELL Technologies). Cells were sample-tagged with anti-mouse MHC-H2 class I antibodies and stained for surface markers. Live CD138- B220+ CD45.1+ CD45.2- cells were purified by FACS sorting. Post-sorting, cell viability was confirmed using a Cellometer K2 Image Cytometer. Pooled single-cell suspensions were loaded into a BD Rhapsody cartridge targeting 100,000 cells per lane. Libraries were prepared following the BD Rhapsody System Mouse TCR/BCR, Targeted mRNA, and Sample Tag Library Preparation Protocol. Library QC was performed using an Agilent 5200 Fragment Analyzer and quantified via Qubit. Sequencing was performed on an Element Biosciences Aviti System using a High Output 2x75 cycles Flowcell.