Using LUMA: a Luminometric-based assay for global DNA-methylation.
Changes in genomic DNA methylation are important events in normal and pathological cellular processes, contributing both to normal development and differentiation as well as to cancer and other diseases. We describe here a method to analyze global genomic DNA methylation, using a luminometric technology to quantitate methylation sensitive restriction digestions. The method is called LUminometric Methylation Assay (LUMA), and is based on a polymerase extension assay using the Pyrosequencing platform. The method is quantitative, highly reproducible and uses an internal control for DNA input. No modification of genomic DNA is needed and the total running time is only six hours. The method is suitable for analyzing clinical material, as well as determining dynamic changes in global methylation/demethylation events. This report describes the method in detail and gives an example of its application in epigenetic research.
Karimi
Mohsen
M
Department of Clinical Neuroscience, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden.
Johansson
Sofia
S
Ekström
Tomas J
TJ
D000818:Animals; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D016678:Genome / Q000235:genetics*; D006801:Humans; D008163:Luminescent Measurements; D008722:Methods