Skip to main content

Methylation-Sensitive Restriction Fingerprinting

  • Protocol
DNA Methylation Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 200))

Abstract

Methylation of cytosine residues is an almost ubiquitous finding of higher organisms (1). The majority of this methylation occurs at the dinucleotide CpG (where p denotes a phosphate group) (2). CpG sites are distributed throughout the genome with clusters of the sequence being found in the 5′ promoter region of housekeeping genes, in groups known as CpG islands. These short stretches of DNA have a have a C and G base composition, which in mammals and avians is estimated to be 10 times higher than in bulk DNA (3)

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 69.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Bestor, T. H. (1990) DNA methylation: evolution of a bacterial immuno function into a regulator of gene expression and genome structure in higher eukaryotes. Phil. Trans. R. Soc. Lond. B 326x326, 179–187.

    Article  Google Scholar 

  2. Gruenbaum, Y., Stein, R., Cedar, H., and Razin, A. (1981) Methylation of CpG sequences in eukaryotic DNA. FEBS Lett. 124, 67–71.

    Article  PubMed  CAS  Google Scholar 

  3. Bird, A. P. (1986) CpG rich islands and the functions of DNA methylation. Nature 321, 209–213.

    Article  PubMed  CAS  Google Scholar 

  4. Riggs, A. D. and Pfeifer, G. P. (1992) X chromosome inactivation and cell memory. Trends Genet. 8, 169–174.

    PubMed  CAS  Google Scholar 

  5. Brandeis, M., Frank, D., Keshet, I., Seigfried, Z., Mendelsohn, M., Nemes, A., et al. (1994) Sp1 elements protect a CpG island from de novo methylation. Nature 371, 435–438.

    Article  PubMed  CAS  Google Scholar 

  6. Stein, R., Sciaky-Gallili, N, Razin, A., and Cedar, H. (1982) Pattern of methylation of two genes coding for housekeeping gene functions. Proc. Natl. Acad. Sci. USA 79, 2422–2426.

    Google Scholar 

  7. Issa, J.-P. J., Vertino, P. M., Wu, J., Sazawal, S., Celano, P., Nelkin, B. D., et al. (1993) Increased cytosine DNA-methyltransferase activity during colon cancer progression. J. Natl. Canc. Inst. 85, 1235–1240.

    Article  CAS  Google Scholar 

  8. Melki, J. R., Warnecke, P., Vincent, P. C., and Clark, S. J. (1998) Increased DNA methyltransferase expression in leukaemia. Leukaemia 12, 311–316.

    Article  CAS  Google Scholar 

  9. Herman, J. G., Jen, J., Merlo, A., and Baylin, S. B. (1996) Hypermethylation-associated inactivation indicates a tumour supressor role for p15INK4B. Cancer Res. 56, 722–727.

    PubMed  CAS  Google Scholar 

  10. Huang, T. H. M., Laux, D. E., Hamlin, B. C., Tran, P., Tran, H., and Lubahn, D. B. (1997) Identification of DNA methylation markers for human breast carcinomas using the methylation-sensitive restriction fingerprint technique. Cancer Res 57, 1030–1034.

    PubMed  CAS  Google Scholar 

  11. Southern, E. M. (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98, 503–506.

    Article  PubMed  CAS  Google Scholar 

  12. Zeng, M., Martsen, E. O., and Lapeyre, J. N. (1998) Re-amplification of short primer-generated bands from RAPD and methylation-sensitive restriction fingerprinting by discrimination primers.Biotechniques 24, 402–403.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2002 Humana Press Inc.

About this protocol

Cite this protocol

Davies, C.S. (2002). Methylation-Sensitive Restriction Fingerprinting. In: Mills, K.I., Ramsahoye, B.H. (eds) DNA Methylation Protocols. Methods in Molecular Biology™, vol 200. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-182-5:043

Download citation

  • DOI: https://doi.org/10.1385/1-59259-182-5:043

  • Publisher Name: Springer, Totowa, NJ

  • Print ISBN: 978-0-89603-618-5

  • Online ISBN: 978-1-59259-182-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics