The NuProbe team has pioneered the development of a unique and novel PCR-based enrichment method called Blocker Displacement Amplification (BDA). The BDA technology enables the selective amplification of low abundant sequence variants (SNV and indels) in a background of wildtype DNA.
Systematic experimental validation of BDA enrichment on hundreds of different SNVs and on a variety of samples ranging from synthetic templates to cell-line genomic DNA to clinical blood samples have demonstrated the robustness of BDA as a rare variant enrichment and detection method.
- Ultrasentive and quantitative: BDA allows for easy detection and quantitation of hundreds of potential variants down to less than 0.1% allele frequency. We have shown that BDA yields a median enrichment greater than 1,000 fold across 200+ SNPs.
- High multiplexing capabilities: BDA has a broad temperature range spanning a 8 °C window (56 – 64 °C) over which it effectively enriches variants. This temperature robustness is a key feature that provides practical advantages for nucleic acid testing by facilitating highly multiplexed enrichment of many SNVs.
- Simple method: BDA requires little to no empirical protocol optimization, employs unmodified and broadly available DNA oligonucleotides, is broadly compatible with many enzymes, uniquely robust to buffer conditions and tolerant to instrument temperature inaccuracies and non-uniformity.