The Use of Immobilized Mismatch Binding Protein in Mutation/SNP Detection

  • Robert Wagner
  • Alan Dean
Part of the Methods in Molecular Biology™ book series (MIMB, volume 152)


The detection of single-base change mutations and polymorphisms is of enormous importance, both in research and in diagnostics. The ability to identify and score single nucleotide polymorphisms (SNPs) is becoming a key element of gene identification and mapping, and the future of human diagnostics depends on having the ability to detect single-base change mutations, because these represent the vast majority of disease-causing and diseaseassociated mutations. An ideal system for detecting and scoring these mutations and SNPs will have certain key characteristics: (1) robustness: the method will be user friendly and not subject to wide fluctuations caused by small changes in experimental conditions; (2) high throughput: given the requirements of genomic research and large-scale diagnostics, a useful method of mutation/SNP detection must be able to handle thousands of samples per day with limited technician effort; (3) low cost: for wide-spread use in both research and clinical diagnostics, low-cost and easy availability of both equipment and reagents is crucial; (4) no gels: this requirement is primarily to meet the high throughput requirement; (5) no radioactivity: given the problems of radioactive material handling and disposal and the availability of a wide variety of alternatives, radioactivity should not be a part of the ideal mutation detection system. Although there may be additional preferences of individual researchers, any mutation/SNP detection system that successfully


Probe Preparation Mismatch Repair System Rare Sequence Unpaired Basis Unlabeled Primer 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Modrich, P. and Lahue, R. (1996) Mismatch repair in replication fidelity, genetic recombination, and cancer biology. Annu. Rev. Biochem. 65, 101–133.PubMedCrossRefGoogle Scholar
  2. 2.
    Dohet, C., Wagner, R., and Radman, M. (1985) Repair of defined single-base-pair mismatches in Escherichia coli. Proc. Natl. Acad. Sci. USA 82, 503–505.PubMedCrossRefGoogle Scholar
  3. 3.
    Su, S.-S., Lahue, R. S., Au, K. G., and Modrich, P. (1988) Mispair specificity of methyl-directed DNA mismatch correction in vitro. J. Biol. Chem. 263, 6829–6835.PubMedGoogle Scholar
  4. 4.
    Fazakerley, G. V., Quignard, E., Woisard, A., et al. (1986) Structures of mismatched basepairs in DNA and their recognition by the E. coli mismatch repair system. EMBO J. 5, 3697–3703.PubMedGoogle Scholar
  5. 5.
    Jones, M., Wagner, R., and Radman, M. (1987) Repair of a mismatch is influenced by the base composition of the surrounding nucleotide sequence. Genetics 115, 605–610.PubMedGoogle Scholar
  6. 6.
    Parker, B. O. and Marinus, M. G. (1992) Repair of DNA heteroduplexes containing small heterologous sequences in Escherichia coli. Proc. Natl. Acad. Sci. USA 89, 1730–1734.PubMedCrossRefGoogle Scholar
  7. 7.
    Wagner, R., Debbie, P., and Radman, M. (1995) Mutation detection using immobilized mismatch binding protein (MutS). Nucleic Acids Res. 23, 3944–3948.PubMedCrossRefGoogle Scholar
  8. 8.
    Debbie, P., Young, K., Pooler, L., et al. (1997) Allele identification using immobilized mismatch binding protein: detection and identification of antibiotic resistant bacteria and determination of sheep susceptibility to scrapie. Nucleic Acids Res. 25, 4825–4829.PubMedCrossRefGoogle Scholar
  9. 9.
    Wagner, R. and Dean, A. (1999) The use of immobilized mismatch binding protein for the optimization of PCR fidelity, in PCR Applications (Innis, M., Gelfand, M., and Sninsky, J., eds.), Academic, New York, pp. 95–104.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2000

Authors and Affiliations

  • Robert Wagner
    • 1
  • Alan Dean
    • 1
  1. 1.Gene Check Inc.Fort Collins

Personalised recommendations