Manage sequence Tm using locked nucleic acid bases
Because of the afforded increase in Tm, locked nucleic acid qPCR probes can be designed with shorter lengths than standard probes. Shorter probes are more effectively quenched and have a higher signal-to-noise ratio. They are, therefore, more
sensitive, providing robust target detection regardless of sequence GC content.
These probes also show greater mismatch discrimination compared to traditional qPCR probes. This improves their ability to distinguish mutations or single nucleotide polymorphisms (SNPs) . By including locked nucleic acid bases, you can design a probe
with ΔTm >15°C. By varying the number of these modified bases, you can essentially control the Tm of a nucleotide duplex. Use these probes in methods that use differential hybridization to distinguish polymorphisms.
Other applications of locked nucleic acid probes include transcript variant identification; verification of microbial species; and improved target detection in FFPE tissue, biofluids, and other challenging samples.