Question: Why isn’t TotalSeq™️-B compatible with Fixed RNA Profiling for Multiplexed Samples?
Answer: Assignment of library molecules to samples relies on a barcode molecule.
Probes are assigned to samples based on the presence of both a unique GEM Barcode and Probe Barcode. Feature Barcode molecules are assigned to samples based on a GEM Barcode and Antibody Barcode.
TotalSeq™️-B Feature Barcode libraries only have a GEM Barcode, and thus cannot be accurately assigned to samples in the context of mutliplets.
Multiplets are more likely to arise if samples are multiplexed. If using TotalSeq™️-B in the context of Multiplexing, this would lead to wasted Feature Barcode reads as sample-specific Antibody Barcodes aren’t present.
Product: Fixed RNA Profiling, Single Cell Gene Expression Flex