C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/68 (2006.01)
Patent
CA 2395047
The invention relates to a method for the parallel detection of the degree of methylation of genomic DNA wherein the following the steps are performed: (a) chemical treatment at the 5' position of non-methylated cytosine bases converts said bases into uracil, thymidine or another base which exhibits hybridization behavior different to that of cytosine in a genomic DNA sample; (b) more than ten different fragments, each having less than 2000 base pairs in said chemically treated genomic DNA sample, are amplified simultaneously using synthetic oligonucleotides as a primer, whereby said primers each contain genomic sequences which are involved in gene regulation and/or transcribed and/or translated, such as those sequences which should be obtained after execution of steps (a); (c) the sequence contexts of all or a portion of the CpG dinucleotides or CpNpG trinucleotides contained in the amplified fragments are determined.
L'invention concerne un procédé de détection parallèle de l'état de méthylation d'ADN génomique, comprenant les étapes suivantes : (a) dans un échantillon d'ADN génomique, on convertit en position 5', par traitement chimique, des bases cytosine non méthylées en uracile, thymidine ou une autre base dont le comportement d'hybridation est différent de celui de la cytosine ; b) on amplifie, dans cet ADN génomique traité chimiquement, plus de dix fragments différents présentant une longueur inférieure à 2000 paires de bases, en utilisant simultanément des oligonucléotides de synthèse comme amorces, ces dernières contenant chacune des séquences génomiques participant à au contrôle de l'expression génique et/ou transcrites et/ou traduites, similaires à celles qui seraient obtenues après le traitement de l'étape a) ; c) on détermine le contexte des séquences de la totalité ou d'une partie des CpG dinucléotides ou trinucléotides CpNpG contenus dans les fragments amplifiés.
Olek Alexander
Piepenbrock Christian
Epigenomics Ag
Macrae & Co.
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