Locked Nucleic Acid Thymine Monomer Probe Identifies Four Single-Nucleotide Variants by Melting Temperature

High-resolution melting (HRM) analysis of DNA is a closed-tube SNP detection method that has shown many advantages in clinical laboratory, point-of-care diagnostics, and personalized medicine. While recently developed melting probes have significantly improved the discrimination of mismatched (mutant) alleles from matched (wild-type) alleles, no one attempts to design a simple probe that can reliably distinguish mismatched alleles among themselves. Such a new probe could facilitate the discovery of rare genetic mutations at lower cost.