Restriction Digestion and Real-Time PCR (qAMP)

  • Christopher C. Oakes
  • Sophie La Salle
  • Jacquetta M. Trasler
  • Bernard Robaire
Part of the Methods in Molecular Biology book series (MIMB, volume 507)

Abstract

DNA methylation in mammals has been shown to play many important roles in diverse biological phenomena. Here we describe a simple and straightforward method that quantitatively measures site-specific levels of DNA methylation in a quick and cost-effective manner. The quantitative analysis of DNA methylation using real-time PCR (qAMP) technique involves the digestion of genomic DNA using methylation-sensitive and methylation-dependent restriction enzymes followed by real-time PCR. This approach generates accurate and reproducible results without the requirement for prior treatment of the DNA with sodium bisulfite.

Keywords

DNA methylation analysis method restriction enzyme real-time PCR 

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Christopher C. Oakes
    • 1
  • Sophie La Salle
    • 1
  • Jacquetta M. Trasler
    • 2
  • Bernard Robaire
    • 3
  1. 1.Department of Pharmacology and Therapeutics and The Montreal Children’s Hospital Research InstituteMcGill UniversityQuebecCanada
  2. 2.Department of Pharmacology and Therapeutics, Department of Pediatrics, Department of Human Genetics and The Montreal Children’s Hospital Research InstituteMcGill UniversityQuebecCanada
  3. 3.Department of Pharmacology and Therapeutics and Department of Obstetrics and GynecologyMcGill UniversityQuebecCanada

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