This dataset was generated by using Gene Expression Flex with Feature Barcode and Proteintech Genomics MultiPro® Human Discovery Panel (Catalog #G900150) to obtain transcriptome and proteome information.
DTCs and matched PBMCs were obtained from Discovery Life Sciences, including Stage I kidney cancer DTCs, Stage I kidney cancer matched PBMCs, Stage III kidney cancer DTCs, Stage III kidney cancer matched PBMCs, Stage III lung cancer DTCs, and Stage III lung cancer matched PBMCs. Control PBMCs were obtained from AllCells.
Samples were prepared in accordance with the Cell Preparation Guide Handbook (CG00053) for Tips & Best Practices on handling and counting cells. Consult Cell Thawing Protocols for Single Cell Assays (CG000447) for guidance on thawing dissociated tumor cells.
Antibody staining was performed as described in Intracellular Protein Labeling Protocol section of the Demonstrated Protocol Cell Surface & Intracellular Protein Labeling for GEM-X Flex Gene Expression (CG000781, Rev A) and Flex assay was performed as described in the GEM-X Flex Gene Expression Reagent Kits for Multiplexed Samples with Feature Barcode technology for Protein using Barcode Oligo Capture User Guide (CG000789). Libraries were sequenced on an Illumina NovaSeq 6000 with approximately 40,000 paired end reads/cell for Gene Expression and 20,000 paired end reads/cell for Feature Barcode libraries.
Dual indexing libraries were sequenced with the following scheme:
- 28 cycles Read 1
- 10 cycles i7
- 10 cycles i5
- 90 cycles Read 2
Number of cells called by condition: PBMC_Control1: 20,490 PBMC_Control2: 18,232 Kidney_Cancer_DTC: 23,574 Kidney_Cancer_PBMC: 20,066 Kidney_Sarcomatoid_DTC: 21,052 Kidney_Sarcomatoid_PBMC: 18,344 Lung_Cancer_DTC: 20,423 Lung_Cancer_PBMC: 15,725