Structure-specific DNA cleavage on surfaces

J Am Chem Soc. 2002 Jul 10;124(27):7924-31. doi: 10.1021/ja012082c.

Abstract

The structure-specific invasive cleavage reaction is a useful means for sensitive and specific detection of single nucleotide polymorphisms, or SNPs, directly from genomic DNA without a need for prior target amplification. A new approach integrating this invasive cleavage assay and surface DNA array technology has been developed for potentially large-scale SNP scoring in a parallel format. Two surface invasive cleavage reaction strategies were designed and implemented for a model SNP system in codon 158 of the human ApoE gene. The upstream oligonucleotide, which is required for the invasive cleavage reaction, is either co-immobilized on the surface along with the probe oligonucleotide or alternatively added in solution. The ability of this approach to unambiguously discriminate a single base difference was demonstrated using PCR-amplified human genomic DNA. A theoretical model relating the surface fluorescence intensity to the progress of the invasive cleavage reaction was developed and agreed well with experimental results.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • DNA / chemistry*
  • DNA / genetics
  • DNA / metabolism
  • Genome, Human
  • Humans
  • Models, Genetic
  • Polymorphism, Single Nucleotide
  • Substrate Specificity
  • Surface Properties
  • Temperature

Substances

  • DNA