Simple SNP typing assay using a base-discriminating fluorescent probe

Mol Biosyst. 2006 Feb;2(2):122-7. doi: 10.1039/b515923g. Epub 2006 Jan 5.

Abstract

We have developed a new concept involving a single-step homogeneous method for single-nucleotide polymorphism (SNP) typing. In this method, a probe containing base-discriminating fluorescent (BDF) bases is added to a sample solution. BDF base-containing DNA usually shows only a weak fluorescence, but emits a strong blue fluorescence when it recognizes a target base at a specific site in a hybridized strand. By utilizing this feature, a simple mix-and-read SNP typing assay was achieved without any tedious probe-designing or washing processes for exclusion of hybridization error or any addition of DNA-modifying enzymes. This is very different from conventional methods. We simultaneously analyzed a number of samples with ease, with a high accuracy, using our BDF assay.

MeSH terms

  • Base Sequence
  • Biological Assay*
  • DNA / chemistry*
  • DNA Probes / chemical synthesis*
  • DNA Probes / chemistry
  • Female
  • Fluorescent Dyes / chemical synthesis*
  • Fluorescent Dyes / chemistry
  • Humans
  • Models, Molecular
  • Nucleic Acid Hybridization
  • Polymerase Chain Reaction
  • Polymorphism, Single Nucleotide*
  • Spectrometry, Fluorescence

Substances

  • DNA Probes
  • Fluorescent Dyes
  • DNA