Question: What is the difference between TotalSeq A, B, and C?
Answer: BioLegend offers three different formats of antibody-oligonucleotide conjugates as part of their TotalSeq line. These antibody formats are TotalSeq A, TotalSeq B, and TotalSeq C. Each format has a different sequence used for capturing the antibody barcode during library construction.
- TotalSeq A is compatible with any Single Cell RNA-seq product that uses poly(dT) for mRNA capture. This format can be used for CITE-seq and REAP-seq applications. TotalSeq A is not supported by 10x Genomics. Please contact New York Genome Center or BioLegend for assistance with the assay or software.
- TotalSeq B is specifically designed to be compatible with the Single Cell 3’ v3 workflow and is fully supported by 10x Genomics. It contains a capture sequence ("Capture Sequence 1") complementary to the capture sequence on the Single Cell 3' v3 Gel Bead oligos. To generate Cell Surface Protein libraries from TotalSeq B antibodies, please see chromium single cell user-guide v3 chemistry with feature barcoding technology for cell surface protein
- TotalSeq C is specifically designed to be compatible with the Single Cell 5' workflow and is fully supported by 10x Genomics. It contains a capture sequence that is complementary to the Template Switch Oligo (TSO) on the Single Cell 5' Gel Bead oligos. To generate Cell Surface Protein libraries from TotalSeq C antibodies, please see Chromium Single Cell V(D)J Reagent Kits with Feature Barcoding technology for Cell Surface Protein.
Note: Cell Ranger 3.0 or higher and Loupe Cell Browser 3.0 or higher are required for analyzing Cell Surface Protein libraries.
Products: Single Cell Gene Expression, Single Cell Immune Profiling