Rationally Designed Universal Melting Probes (Uni-Melt) for Multiplex Genotyping

copies/μL. In addition, Uni-Melt can differentiate multiple mutation sites including single nucleotide polymorphisms (SNPs), as demonstrated in the SLC25A13 gene. Overall, Uni-Melt is a closed-tube, sensitive, and specific system for multiplexed genotyping compatible with commonly employed quantitative polymerase chain reaction (qPCR) thermocyclers. Compared to the traditional probe-based melting curve analysis, Uni-Melt can reduce the assay cost associated with multiple probe modifications and decrease the complexity of the reaction without sacrificing sensitivity and specificity, which makes it a practically useful tool for clinical screening.