Conditionally fluorescent molecular probes for detecting single base changes in double-stranded DNA.

ABSTRACT: Small variations in nucleic acid sequences can have far-reaching phenotypic consequences. Reliably distinguishing closely related sequences is therefore important for research and clinical applications. Here, we demonstrate that conditionally fluorescent DNA probes are capable of distinguishing variations of a single base in a stretch of target DNA. These probes use a novel programmable mechanism in which each single nucleotide polymorphism generates two thermodynamically destabilizing mismatch bubbles rather than the single mismatch formed during typical hybridization-based assays. Up to a 12,000-fold excess of a target that contains a single nucleotide polymorphism is required to generate the same fluorescence as one equivalent of the intended target, and detection works reliably over a

SUBMITTER: Chen SX 

PROVIDER: S-EPMC3844531 | biostudies-literature | 2013 Sep

REPOSITORIES: biostudies-literature