Human peripheral blood mononuclear cells (PBMCs) of a healthy male donor aged 30-35 were obtained by 10x Genomics from AllCells. Nuclei were isolated using the demonstrated protocol Nuclei Isolation for Single Cell Multiome ATAC + Gene Expression Sequencing (CG000365).
Gene Expression and ATAC libraries were generated from ~16,100 transposed nuclei (10,699 nuclei recovered) as described in the Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Kits User Guide (CG000338 Rev E) using the Chromium Controller and sequenced on an Illumina NovaSeq 6000.
ATAC libraries were sequenced to ~55k read pairs per cell.
- 50 cycles Read 1
- 8 cycles i7 (sample index)
- 24 cycles i5 (10x barcode)
- 49 cycles Read 2.
Gene Expression libraries were sequenced to ~55k reads per cell.
- 28 cycles Read 1 (16 bp barcode, 12 bp UMI)
- 10 cycles i7 (sample index)
- 10 cycles i5 (sample index)
- 90 cycles Read 2 (transcript).