Single-Molecule Counting of Point Mutations by Transient DNA Binding.

ABSTRACT: High-confidence detection of point mutations is important for disease diagnosis and clinical practice. Hybridization probes are extensively used, but are hindered by their poor single-nucleotide selectivity. Shortening the length of DNA hybridization probes weakens the stability of the probe-target duplex, leading to transient binding between complementary sequences. The kinetics of probe-target binding events are highly dependent on the number of complementary base pairs. Here, we present a single-molecule assay for point mutation detection based on transient DNA binding and use of total internal reflection fluorescence microscopy. Statistical analysis of single-molecule kinetics enabled us to effectively discriminate between wild type DNA sequences and single-nucleotide variants at the s

SUBMITTER: Su X 

PROVIDER: S-EPMC5338343 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature