Haplotype of multiple polymorphisms resolved by enzymatic amplification of single DNA molecules

Proc Natl Acad Sci U S A. 1990 Aug;87(16):6296-300. doi: 10.1073/pnas.87.16.6296.


We have developed a reliable method for the direct resolution of haplotypes or linkage phase from individuals who are multiply heterozygous in a given genomic region. The method is based on single-molecule dilution (SMD) of genomic template and amplification via biphasic polymerase chain reaction (booster PCR). We have verified the feasibility of the SMD method for a highly polymorphic region within the beta-globin cluster by analysis of triply heterozygous individuals of known haplotype. This approach should be useful in many studies in population or evolutionary genetics and in a variety of clinical settings.

Publication types

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

MeSH terms

  • Base Sequence
  • DNA / genetics*
  • Deoxyribonucleases, Type II Site-Specific
  • European Continental Ancestry Group / genetics
  • Genetic Carrier Screening
  • Genetic Linkage
  • Haplotypes*
  • Humans
  • Molecular Sequence Data
  • Nucleic Acid Amplification Techniques*
  • Oligonucleotide Probes
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Genetic*
  • Polymorphism, Restriction Fragment Length*
  • Tourette Syndrome / genetics


  • Oligonucleotide Probes
  • DNA
  • Deoxyribonucleases, Type II Site-Specific
  • TCGA-specific type II deoxyribonucleases