Notl-Msel methylation-sensitive amplified fragment length polymorphism for DNA methylation analysis of human cancers

Fumiichiro Yamamoto, Miyako Yamamoto, Jose Luis Soto, Eijiro Kojima, Emily N. Wang, Manuel Perucho, Takao Sekiya, Hiroyasu Yamanaka

Resultado de la investigación: Contribución a una revistaArtículorevisión exhaustiva

31 Citas (Scopus)

Resumen

We have applied a methylation-sensitive restriction endonuclease, Notl, to the existing amplified fragment length polymorphism (AFLP) method and developed Notl-Msel methylation-sensitive-AFLP (MS-AFLP). Notl-Msel MS-AFLP allows the analysis of DNA methylation alterations at the Notl sites scattered over the genome. Hypermethylation and hypomethylation are visualized by the decrease and increase in the band intensity of DNA fingerprints. Identification of consistent changes can be facilitated through parallel electrophoresis of multiple samples. DNA fragments exhibiting alterations can be cloned from fingerprint bands by amplification of gel-eluted DNA with the same pair of primers used for radioactive fingerprint presentation. Fluorescent Notl-Msel MS-AFLP offers a safer method of studying the alterations in DNA methylation, and may be applied to the hybridization of DNA microarrays in the future. Using Notl-Msel MS-AFLP, we observed frequent hypomethylation of a satellite DNA repeat sequence in a majority of breast tumors.

Idioma originalInglés
Páginas (desde-hasta)1946-1956
Número de páginas11
PublicaciónElectrophoresis
Volumen22
N.º10
DOI
EstadoPublicada - 2001

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