LightSNIP - Typing Assays for LightCycler Instruments
Single Nucleotide Polymorphisms (SNP) are used to map genes and to correlate gene loci with diseases, predispositions and even bahaviour. Genetic instructions run hundret thousands of samples to find correlations by statistical methods. In the past the mutation analysis os SNP analysis has been analyzed by PCR based Restriction Fragment Length Polymorphism (RFLP), allele specific oligonucleotides (ASO), DNA sequencing, or solid phase based methods (DNA arrays, chips, beads). The more advanced Real-Time PCR based methods deliver results directly from the reaction mix, avoiding hands on time, reducing the risk of contaminations and reducing the time af analysis.
The two principal methods currently applied for SNP analysis in Real-Time PCR are Hydrolosis (TaqMan) probe assays and probe melting analysis. The former requires a pair of competitive specific probes; one for each variant. There are several disadvantages to this approach. Each SNP requires two detection channels, limiting the potential throughput. Any cross reactivity of the probes with the unspecific target has to be excluded. But the most crucial factor is the influence of additional sequence variations within the probe targeting region that can lead to a signal loss for one allele falsely suggesting a homozygous genetic background.
In contrast, probe melting analysis based assays provide a greater insight into the target sequence, reporting exact temperatures for both genotypes and distinct different curves for new variants. The best estabilished binding probes are the FRET energy transfer based hybridization probes, which use a paior of probes with a donor and an acceptor dye. Other estabilished single probe systems include Molecular Beacons, LightUp probes and SimpleProbe probes.
LightSNIP assays are compatible with all LightCycler models
All LightSNIP assays are tested on the LightCycler 480 instrument.Pur LightSNIP typing assays are also compatible with the LightTyper high-throughput mutation analysis instrument from Roche.
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A license under U.S. Patents 4,683,202, 4,683,195 and 4,965,188 or their foreign counterparts, owned by Hoffmenn-La Roche Inc. and F. Hoffmann-La Roche Ltd (Roche), has an up-front free component and a running-royalty component. The purchase price of this product ioncludes limited, nontransferable rights under the running-royalty component to use onlu this amount of the product to practice the Polymeraze Chain Reaction (PCR) and related processes described in said patents solely for the research and development activities of the purcharser whis this product is used in conjuction with a thermal cycler whose use is covered by the up-front fee component. Rights to the up-front fee component must be obtained by the end user in order to have a complete licence. These rights under the up-front fee component may be purchased from Perkin-Elmer or obtained by purchasing an authorized thermal cycler. No right to perform or offer commercial services of any kind using PCR, including without limitation reporting the results of purchaser's activities for a fee or other commercial consideration, is hereby granted by implication or estoppel. Further information on purchasing licences to practice the PCR process for research applications may be obtained by contacting the Director ofLicensing at The Perkin-Elmer Corporation, 850 Licoln Center Drive, Foster City, California 94404 or at Roche Molecular Systems Inc. 1145 Atlantic Avenue, Alameda, California 94501. The purchase of this product does not convey any right for its use in clinical diagnostic applications, No rights for TaqMan technology under U.S. Patents 5,210,015 and 5,487,972 are hereby conveyed.