GSM7951847: LN_2_BCR; Mus musculus; RNA-Seq
Bone marrow, lymph node and peripheral blood cells isolated from C57BL/6J female mice were stained with Live/Dead dye and lymphocytes were isolated using a BDFACSAria™ IIISorter. Cells were manually counted by Trypan blue and AO-PI after each centrifugation and resuspension. By using Chromium Next GEM Single Cell 5' Kit v2 (10x Genomics, 1000263) and Chromium Next GEM Chip K Single Cell Kit (10x Genomics, 1000287), we performed single cell TCR/BCR-seq and 5' gene expression profiling. The cell suspension was loaded onto the Chromium single cell controller (10x Genomics) to generate single-cell gel beads in the emulsion according to the manufacturer's protocol. Captured cells were lysed and the released RNA were barcoded through reverse transcription in individual GEMs. Cell-barcoded 5' gene expression libraries and V(D)J enriched TCR/BCR libraries were sequenced on an Illumina NovaSeq6000 system.