A new approach for point mutation detection based on a ligase chain reaction

J Biochem Biophys Methods. 2001 Dec 4;50(1):79-89. doi: 10.1016/s0165-022x(01)00178-6.

Abstract

A new method for the identification of point mutations is proposed. The method is based on ligase chain reaction (LCR) and it includes a procedure for correction of ligation by Cleavase. Reaction products are detected by a colorimetric method after adsorption of the resulting DNA duplexes to the solid phase. One strand of LCR products carries biotin to be bound on a streptavidin-coated microwell. Another strand contains a single-stranded region that is to be coupled with an oligonucleotide carrying a substrate for colorimetric detection. The suggested method has two advantages: (i) use of Cleavase increases the accuracy of ligation and (ii) a template independent ligation does not occur in LCR due to a special design of primers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Biotin / metabolism
  • DNA Ligases / metabolism*
  • DNA Mutational Analysis*
  • Electrophoresis, Polyacrylamide Gel
  • Endodeoxyribonucleases / metabolism*
  • Genetic Techniques*
  • Molecular Sequence Data
  • Mutation
  • Oligonucleotides / chemistry
  • Point Mutation*
  • Sequence Homology, Nucleic Acid

Substances

  • Oligonucleotides
  • Biotin
  • Endodeoxyribonucleases
  • cleavase
  • DNA Ligases