GSM7991691: MedLN; Mus musculus; RNA-Seq
Transferred GFP+ B1-8hi B cells were sorted into a cooled 15 mL tube with 0.04% BSA in PBS using a Sony SH800 cell sorter. 12,000 sorted cells suspension was loaded onto Next GEM Chip G targeting 8000 cells and then ran on a Chromium Controller instrument to generate GEM emulsion. Single-cell 3' RNA-seq libraries were generated according to the manufacturer's protocol (10x Genomics Chromium Single Cell 3' Reagent Kit User Guide v3/v3.1 Chemistry). Final libraries were quantified using NEBNext Library Quant Kit for Illumina (NEB) and high sensitivity D1000 TapeStation (Agilent). Libraries were pooled according to targeted cell number, aiming for ~20,000 reads per cell. Pooled libraries were sequenced on a NovaSeq 6000 instrument using an SP 100 cycles reagent kit (Illumina).