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Protocols

 

Cell Labeling:

 

1. Single Cell Labeling with the BD™ Single-Cell Multiplexing Kits:

 

The BD™ Human Single-Cell Multiplexing Kit (Cat. No. 633781) and the BD™ Mouse Immune Single-Cell Multiplexing Kit (Cat. No. 633793) utilize an innovative antibody-oligo technology to provide higher sample throughput for single cell library preparation. Every antibody-oligo in the BD™ Human Single-Cell Multiplexing Kit, referred to as a Sample Tag, has a unique sample oligo barcode conjugated to a human universal antibody and every Sample Tag in the BD™ Mouse Immune Single-Cell Multiplexing Kit is conjugated to an Anti-Mouse CD45, Clone 30-F11 antibody. Up to 12 samples can be labelled and pooled prior to single cell capture with the BD Rhapsody™ Single-Cell Analysis system.

 

2. Single Cell Labeling with the BD™ Single-Cell Multiplexing Kit and BD™ AbSeq Ab-Oligos

 

This protocol describes the use of BD AbSeq Ab-Oligos (antibody-oligonucleotides) with the BD™ Human Single-Cell Multiplexing Kit (Cat. No. 633781) or BD™ Mouse Immune Single-Cell Multiplexing Kit (Cat. No. 633793). The BD™ AbSeq Ab-Oligos are used for antigen-expression profiling with BD Rhapsody™ single cell capture and downstream library preparation. Each BD™ AbSeq Ab-Oligo is an oligonucleotide-conjugated antibody that contains an antibody-specific barcode and poly(A)-tail for bead capture, PCR amplification, and library generation. The protocol supports BD™ AbSeq labeling of 20,000 to 1 million cells. Up to 40 antibodies can be pooled together per staining reaction. The BD Single-Cell Multiplexing Kits utilize an innovative antibody-oligo technology to provide higher sample throughput for single cell library preparation. Every antibody-oligo in the BD™ Human Single-Cell Multiplexing Kit, referred to as a Sample Tag, has a unique sample oligo barcode conjugated to a human universal antibody and every Sample Tag in the BD™ Mouse Immune Single-Cell Multiplexing Kit is conjugated to an Anti-Mouse CD45, Clone 30-F11 antibody. Up to 12 samples can be labelled and pooled prior to single cell capture with the BD Rhapsody™ Single-Cell Analysis system. You can co-label cells with Sample Tags and BD AbSeq Ab-Oligos in a single tube, or you can sequentially label cells with Sample Tags and pool cells before labeling with BD AbSeq Ab-Oligos.