Method for identifying cytosine methylation patterns in...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2351143

The invention relates to a method for identifying 5-methylcytosine positions in genomic DNA. The method is characterised by the following steps: a) the genomic DNA of a cell, a cell line, a tissue or an individual is chemically treated in such a way that the cytosine and 5-methylcytosine react differently and the two products present different base pairing behaviour in the duplex; b) the same nucleic acid section is amplified by means of a polymerase reaction; c) the same nucleic acid section of at least one other cell, cell line, tissue or individual or of any given reference DNA is treated according to steps a) and b); d) heteroduplexes are formed from the at least two amplification products obtained in steps b) and c); and e) a detectable marking is introduced into the heteroduplexes by means of a reaction that is specific to non-complementary base pairs.

L'invention concerne un procédé d'identification de positions 5-Méthylcytosine dans l'ADN génomique. Ce procédé consiste a) à traiter chimiquement l'ADN génomique d'une cellule, d'une lignée cellulaire, d'un tissu ou d'un individu de telle façon que la cytosine et la 5-méthylcytosine réagissent différemment et que l'on obtienne dans les deux produits un comportement d'appariement des bases différent dans la double hélice; b) à amplifier la section d'acide nucléique de ce dernier par une réaction de polymérisation; c) à traiter la même section d'acide nucléique au moins d'une autre cellule, lignée cellulaire, d'un autre tissu ou individu ou d'un ADN de référence quelconque selon a) et b); d) à former des doubles hélices à partir d'au moins deux produits d'amplification obtenus en b) et c); e) à introduire un marquage repérable dans l'hétéroduplex par une réaction spécifique à des paires de bases non complémentaires.

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