Restriction digestion and real-time PCR (qAMP).
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.
Oakes
Christopher C
CC
Department of Pharmacology and Therapeutics and The Montreal Children's Hospital Research Institute, McGill University, Quebec, Montreal, Canada.
La Salle
Sophie
S
Trasler
Jacquetta M
JM
Robaire
Bernard
B
D000818:Animals; D018899:CpG Islands; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D019175:DNA Methylation; D017931:DNA Primers; D004262:DNA Restriction Enzymes; D008297:Male; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D004247:DNA; D004262:DNA Restriction Enzymes