The Tapestri platform supports sample multiplexing using Genotype or Antibody Hashing.
Genotype Hashing (GT)
Genotype hashing uses the differences in germline variants across samples for demultiplexing. Each sample requires a minimum of 5 germline SNPs that distinguish the sample from other samples in the same Tapestri run, though >10 variants are recommended. Generally ~5 germline SNPs can be found per 100 amplicons in Mission Bio DNA panels. When designing panels <200 amplicons, Reach out to support@missionbio.com. It may be advisable to include ~30 amplicons specifically targeting high minor allele frequency SNPs in the genome (these can be drawn from the Mission Bio wgCNV panel or found using dbSNP).
While germline SNP information may be obtained from WGS/WES/orthogonal NGS data, we highly recommend using the Tapestri Bulk DNA Sequencing Kit to identify high quality SNPs for use in demultiplexing. The bulk kit can be run alongside Tapestri runs and the libraries can be processed on the Tapestri Pipeline to automate suitable germline SNP identification.
Antibody Hashing (Ab)
Antibody hashing uses labeled antibodies against CD298 and β2 microglobulin for demultiplexing. Hashtag antibodies should be purchased directly from BioLegend. Each sample must be stained separately and then mixed prior to the Tapestri run. Antibody hashing requires the use of the Mission Bio Protein Staining Kit.
Comparing Genotype and Antibody Hashing
| Genotype Hashing | Antibody Hashing | |
| Sample types | Intact cells, nuclei, low viability samples | High viability intact cells |
| Species input | Any supported species | Human and mouse only |
| Sample Requirements | Requires unique germline SNPs (different patients) | Can multiplex the same patient in one run (ie multiple time points) |
| Cell input requirements | >250,000 total | >500,000, >70% viability per sample |
| Hands on time | Supports DNA only workflow | Requires DNA + Protein workflow |
| DNA Panel | Requires germline SNPs (10+ recommended) | Any DNA panel |