Question: What is the difference between TotalSeq A, B, and C?
Answer: BioLegend offers three different formats of antibody-oligonucleotide conjugates as part of its 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. As this product requires reagent and protocol modifications, 10x Genomics provides only limited support for TotalSeq A. Please contact New York Genome Center or BioLegend for additional assistance with the assay or software.
- TotalSeq B is specifically designed to be compatible with the Single Cell 3’ v3 and 3.1 workflows and is fully supported by 10x Genomics. TotalSeq B antibodies are also compatible with the Fixed RNA Profiling workflow. 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-Guides 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 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 hashing using TotalSeq B or TotalSeq C is not officially supported by 10x Genomics, more details can be found in Can I perform Cell Hashing in the 5' workflow? 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, Fixed RNA Profiling Gene Expression