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  1. Stain with TotalSeq™ reagents
  2. Run single cell platform, with Antibody- Derived Tags (ADT, antibody-oligo conjugates) amplification
  3. Separate ADT-derived cDNA from mRNA-derived cDNA
  4. Amplify both cDNA libraries
  5. Sequence libraries

 

Fig 1. Diagram illustrating CITE-seq workflow. Step 2 can be performed by Drop-seq (depicted in the figure), or another compatible method such as 10X Genomics platform.

 

 

Fig 2. Clustering of approximately 5,000 CITE-seq single-cell expression profiles of PBMCs reveals distinct cell populations based on transcriptome analysis. The left panel shows a two-dimensional representation (tSNE) of global gene expression relationships among all cells. Major cell types in peripheral blood can be discerned based on marker gene expression as indicated. The right panels show mRNA (blue) and corresponding Antibody-Derived Tag (ADT, green) signal for the CITE-seq antibody panel projected on the tSNE plot. Darker shading corresponds to higher levels measured.