Affinity Plus locked nucleic acid bases should be placed at the SNP site and on each of the adjacent bases.We recommend using up to 6 Affinity Plus locked nucleic acid bases in an Affinity Plus qPCR Probe.Use the OligoAnalyzer™ Tool to check secondary Therefore, it is important to examine the probe sequence for self-dimer and hairpin formationĪnd minimize designs that allow Affinity Plus:Affinity Plus pairing. The relative binding affinity (T m) of Affinity Plus locked nucleic acid bases are Affinity Plus:Affinity Plus > Affinity Plus:DNA > DNA:DNA.Multiple locked nucleic acid additions can result in an Affinity Plus qPCR Probe with a ΔT m of >15☌. Depending on sequence context, each locked nucleic acid base insertion into a DNA oligo will increase the T m by 3–6☌.An Affinity Plus qPCR Probe can be designed with several locked nucleic acid monomers resulting in a ΔT m of >15☌, which greatly increasesĪccuracy of allele determination in real-time PCR or other methods that use differential hybridization to distinguish polymorphisms. More importantly, locked nucleic acid probes offer an improved ability to distinguish mutations or single nucleotide Shorter probes have better quenching and a higher signal-to-noise ratio and are, therefore, more sensitive. Because locked nucleic acid bases significantly increase T m, Affinity Plus qPCR Probes can be designed with shorter lengths Locked nucleic acid bases, such as Affinity Plus monomers, can be incorporated into effective qPCR probes. Yakima Yellow is a registered trademark of EliTech Group, and Cy is a registered trademark of Cytiva.Īdvantages of Affinity Plus qPCR Probes containing locked nucleic acid modifications Choose Mini Affinity Plus qPCR Probes for digital PCR (dPCR) platforms.Make flexible T m adjustments more easily than possible with MGB probes.Benefit from this affordable approach to increasing assay hybridization specificity.
Improve mismatch discrimination compared to traditional qPCR probes.Select from a wide range of fluorophores including FAM, Yakima Yellow ® (YAK), SUN™, HEX, Cy ® 3, Cy ® 5, TEX, and TYE™ dyes.Probe, locked nucleic acids impart heightened structural stability, leading to increased hybridization melt temperature (T m). (see the technology page, Locked nucleic acids, for more information).
The Affinity Plus bases used in these qPCR probes include up to 6 locked nucleic acid monomers Use Affinity Plus™ qPCR Probes for SNP genotyping, transcript variant identification, and improved target detection in challenging samples (FFPE tissue, biofluids). Increase SNP target specificity and use shorter sequences with these high T m locked nucleic acid probes
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