Question: How many cells should I use as input into labeling and flow sorting in the Barcode Enabled Antigen Mapping (BEAM) workflow?
Answer: The recommended number of starting cells will vary greatly depending on the percentage of T or B cells in the sample, the percentage of those T of B cells that recognize the antigen(s), the sample viability and quality, and the desired number of cells to use as input into the Single Cell 5'v2 assay.
- Start with sufficient cell numbers to account for cell loss at each step of the workflow. This includes cell loss during sample thawing, optional bead-based pre-enrichment, BEAM labeling, washing, and flow sorting. Having at least 1 million cells as input into flow sorting is recommended.
- The BEAM labeling protocol was demonstrated on 0.2 to 8 million cells. If labeling more than 8 million cells, set up a separate labeling reaction in a separate 15 ml tube.
- The volume of BEAM Assembly should be scaled up for every 1 million cells used for labeling. Refer to the BEAM Reagent Assembly, Sample Labeling and Flow Sorting User Guide (CG000595) for further details.
Products: Single Cell Immune Profiling, Barcode Enabled Antigen Mapping (BEAM)