Human peripheral blood mononuclear cells (PBMCs) of a healthy female donor aged 29 were obtained by 10x Genomics from AllCells. The whole transcriptome (parent) library was prepared with Chromium Single Cell V(D)J Reagent Kits User Guide (v1 Chemistry) with Feature Barcode technology for Cell Surface Protein (CG000186). The targeted gene expression libraries were generated with Targeted Gene Expression Reagent Kits User Guide (CG000293) using the Human Immunology Panel (PN-1000246) and sequenced on an Illumina NovaSeq 6000.
Targeted Gene Expression Library
- Sequencing Depth: 10,271 reads per cell
- Paired-end (28 X 91), single indexed Read 1: 26 cycles (16 bp barcode, 10 bp UMI) i7 index: 8 cycles i5 index: 0 cycles Read 2: 91 cycles (transcript)
Note: R1 read requirement is 26bp; actual sequencer configuration was R1=28bp to enable pooling; pipeline trims 2bp from R1.
The key metrics were:
- Number of cells called: 6,805
- Reads Mapped Confidently to Targeted Transcriptome: 75.1%
- Total targeted genes detected: 956