Degenerate oligonucleotide-primed PCR: general amplification of target DNA by a single degenerate primer

Genomics. 1992 Jul;13(3):718-25. doi: 10.1016/0888-7543(92)90147-k.

Abstract

A version of the polymerase chain reaction (PCR), termed degenerate oligonucleotide-primed PCR (DOP-PCR), which employs oligonucleotides of partially degenerate sequence, has been developed for genome mapping studies. This degeneracy, together with a PCR protocol utilizing a low initial annealing temperature, ensures priming from multiple (e.g., approximately 10(6) in human) evenly dispersed sites within a given genome. Furthermore, as efficient amplification is achieved from the genomes of all species tested using the same primer, the method appears to be species-independent. Thus, for the general amplification of target DNA, DOP-PCR has advantages over interspersed repetitive sequence PCR (IRS-PCR), which relies on the appropriate positioning of species-specific repeat elements. In conjunction with chromosome flow sorting, DOP-PCR has been applied to the characterization of abnormal chromosomes and also to the cloning of new markers for specific chromosome regions. DOP-PCR therefore represents a rapid, efficient, and species-independent technique for general DNA amplification.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Chromosome Mapping
  • Cloning, Molecular
  • DNA / genetics*
  • Evaluation Studies as Topic
  • Genetic Markers
  • Humans
  • Mice
  • Molecular Sequence Data
  • Nucleic Acid Amplification Techniques*
  • Oligodeoxyribonucleotides / genetics
  • Polymerase Chain Reaction / methods*

Substances

  • Genetic Markers
  • Oligodeoxyribonucleotides
  • DNA