Enabling genomic technology
Our novel HyBeacon® molecular probes deliver improved differentiation of nucleic acid targets compared to traditional fluorophore-quencher style probes.
Technologies that permit rapid investigation of target nucleic acid sequences, such as those containing single nucleotide polymorphisms (SNPs), insertions, deletions or short tandem repeats (STRs), are of great consequence to genetic analysis. In 2001, LGC developed a novel fluorescent oligonucleotide probe technology, termed HyBeacons. A single HyBeacon probe can be used to differentiate even very closely-related target sequences through end point analysis.
HyBeacons are linear oligonucleotides possessing one or more fluorescent dyes covalently linked to internal nucleotides. The inherent fluorescence-quenching properties of the DNA bases and attached fluorophores cause fluorescence emission to be low in the unstructured single-stranded probe. Hybridisation of HyBeacons to complementary DNA target sequences results in a measurable elevation of probe fluorescence emission. Conversely, when the probe is displaced, the fluorescence is decreased. The internal nature of HyBeacon labelling can be exploited to offer improved differentiation of highly similar DNA sequences during end point analysis using just a single probe.
What are the advantages of using HyBeacon probes for target detection compared to fluorophore-quencher style probes?
- Novel approach – HyBeacon probes are novel meaning they do not infringe the intellectual property that can make other technologies prohibitively expensive
- Reduced cost – a single HyBeacon probe can differentiate multiple sequence variants in the same tube, thereby reducing the number of probes required for reliable allelic discrimination
- Increased capacity – using HyBeacon probes labeled with different dyes permit detection of multiple targets in the same tube, thereby increasing the number of targets detected in the same run
- Reduced handling – HyBeacon master mixes contain DNA polymerases that show significantly improved tolerance to the presence of PCR inhibitors, thereby allowing detection directly from a range of biological samples (e.g. blood, saliva, urine) and eliminating expensive and time-consuming extraction methods
- Simpler design – HyBeacon probes are not restricted by internal sequence requirements (e.g. secondary structure). Furthermore, the number and placement of fluorescent dyes can be optimized to improve the discriminatory characteristics of the probe
- Superior accuracy – HyBeacon probes detect additional unknown polymorphisms / mutations that can result in incorrect calls with alternative probe technologies
- Faster results – sequence variants are detected by end point analysis following a rapid PCR amplification step, which collectively take as little as 45 minutes
- Improved convenience – HyBeacon probes can be labelled with a broad range of fluorophores meaning they are compatible with all commonly-used instruments
HyBeacon Probes Brochure