Chromium single cell 5 library gel bead kit v2

After being counted with C-Chip (inCYTO), CMV-reactivated cells and control cells from all three individuals were mixed separately and diluted with PBS to a final concentration of ~800 cells/μl, and about 20,000 cells per reaction were loaded onto a Chromium Single Cell Chip (10x Genomics). The libraries for RNA-seq and TCR-seq were prepared using the Chromium Single Cell 5′ Library & Gel Bead Kit v2 and Chromium Single Cell V(D)J Human T Cell Enrichment Kit (10x Genomics) following the manufactory’s protocol. Sequences within these libraries were ligated with BGIseq adapters, and then CMV and control libraries were loaded onto the sequencing chip. The RNA-seq libraries were sequenced with an 8-base index read, a 26-base read 1 containing cell-identifying barcodes and unique molecular identifiers (UMIs), and a 100-base read 2 containing transcript sequences on BGIseq500; TCR-seq were sequenced with an 8-base index read, a 150-base read 1 containing cell-identifying barcodes, UMIs and insert starting from the V-gene region, and a 150-base read 2 containing an insert from the C-gene region. The raw data after sequencing were about 10 + 35 Gb per library for RNA-seq and 35 + 35 Gb for TCR-seq.