Detection of DNA methylation in eucaryotic cells.

Anetta Sulewska; Wieslawa Niklinska; Miroslaw Kozlowski; Lukasz Minarowski; Wojciech Naumnik; Jacek Niklinski; Katarzyna Dabrowska; Lech Chyczewski

Vol 45, No 4 (2007)

Original paper

Submitted: 2011-12-19

Published online: 2008-01-01

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Detection of DNA methylation in eucaryotic cells.

Anetta Sulewska, Wieslawa Niklinska, Miroslaw Kozlowski, Lukasz Minarowski, Wojciech Naumnik, Jacek Niklinski, Katarzyna Dabrowska, Lech Chyczewski

Folia Histochem Cytobiol 2007;45(4):315-324.

Vol 45, No 4 (2007)

ORIGINAL PAPERS

Submitted: 2011-12-19

Published online: 2008-01-01

Abstract

The methods of molecular biology allow for analyzing the methylation pattern in the whole genome and in particular genes. We differentiate methylated sequences from unmethylated ones by means of cutting the genomic template with methylation-sensitive restriction enzymes or by sodium bisulfite DNA modification. Chemical modification precedes most quantitative and qualitative PCR techniques: MS-PCR, MS-nested PCR, Real-Time PCR, QAMA, HeavyMethyl, MSHRM. Restriction enzymes, on the other hand, may be used together with PCR or hybridisation methods (Southern blot and microarrays). PCRs are conducted with primers specific for methylated and unmethylated sequences and sometimes, similarly to hybridisation techniques, with specifically labeled probes or dyes intercalating to double-stranded nucleic acids. The most advanced methylation detection techniques (MALDI-TOF MS and HPLC) significantly reduce the amount of biological material used for tests, but they require specialist equipment.