Human Melanoma tumor derived cells of a pre-treatment male donor aged 40 were obtained by 10x Genomics from Discovery Life Sciences.
Gene Expression, BCR Enriched, and TCR Enriched libraries were generated from 16,520 cells (86% viable, expected cell recovery 8,524) as described in the Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index) User Guide (CG000331 Rev A), and sequenced on an Illumina NovaSeq 6000.
The whole transcriptome/gene expression library was also used to generate targeted gene expression libraries as described in the Targeted Gene Expression Reagent Kits User Guide (CG000293) using the Human Pan-Cancer Panel (PN-1000247), Human Immunology Panel (PN-1000246), and the Human Gene Signature Panel (PN-1000245).
Sequencing Configuration:
- Gene Expression: Paired-end (26X90), dual-indexed sequencing. Read 1: 26 cycles, i7 index: 10 cycles, i5 index: 10 cycles, Read 2: 90 cycles
- BCR Enriched: Paired-end (26X90), dual-indexed sequencing. Read 1: 26 cycles, i7 index: 10 cycles, i5 index: 10 cycles, Read 2: 90 cycles
- TCR Enriched: Paired-end (28X90), dual-indexed sequencing. Read 1: 28 cycles, i7 index: 10 cycles, i5 index: 10 cycles, Read 2: 90 cycles. Note: Read 1 requirement is 26 bp; 28 bp Read 1 is trimmed by Cell Ranger pipeline.
Analysis Settings:
- Gene Expression: --expect-cells=10000