title: Detection of DNA methylation in eucaryotic cells.
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.
authors.affiliation: Thoracic Surgery, Medical University of Bialystok. anettasul@poczta.onet.pl
authors.forename: Anetta
authors.initials: A
authors.lastname: Sulewska
authors.forename: Wieslawa
authors.initials: W
authors.lastname: Niklinska
authors.forename: Miroslaw
authors.initials: M
authors.lastname: Kozlowski
authors.forename: Lukasz
authors.initials: L
authors.lastname: Minarowski
authors.forename: Wojciech
authors.initials: W
authors.lastname: Naumnik
authors.forename: Jacek
authors.initials: J
authors.lastname: Niklinski
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Dabrowska
authors.forename: Lech
authors.initials: L
authors.lastname: Chyczewski
mesh_terms: D000818:Animals; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D005057:Eukaryotic Cells / Q000378:metabolism*; D006801:Humans; D046228:Microarray Analysis; D008967:Molecular Biology / Q000379:methods*; D016133:Polymerase Chain Reaction