Process for detecting and quantifying nucleic acid molecules

Details Process for detecting and quantifying nucleic acid molecules Process for detecting and quantifying nucleic acid molecules

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01)

Patent

CA 2241185

The invention relates to a process for detecting and quantifying nucleic acid (=NA) molecules within a population (I) of NA molecules of identical, similar or different sequence, in which: a) heteroduplexes are formed in which nucleotides which are unpaired or, according to Watson and Crick, incorrectly paired are chemically reactive with a carbon di-imide compound; b) the single or double-stranded ribonucleic acid (=RNA) molecules or single-stranded deoxyribonucleic acid (=DNA) molecules contained in population (I) are transferred into a population (II) of double-stranded DNA molecules; c) the population (II) of double-stranded DNA molecules are subjected to denaturation and subsequent renaturation, to produce a mixture of homo and heteroduplex DNA molecules. To make it possible to quantify the NA molecules detected, it is proposed that a purification stage take place between steps a) and b) and between steps b) and c).

L'invention concerne un procédé de détection de et quantification de molécules d'acide nucléique (=AN) dans une population (I) de molécules d'AN de séquence identique, similaire ou différente, selon lequel: a) des hétéroduplex sont formés, dans lesquels des nucléotides non appariés ou appariés de manière impropre selon Watson et Crick, réagissent chimiquement avec un composés carbodi-imide; b) des molécules d'acide ribonucléique (=ARN) à brin unique ou double ou des molécules d'acide desoxyribonucléique (=ADN) à brin unique contenues dans la population (I) sont transférées dans une population (II) de molécules d'ADN à deux brins; c) la population (II) de molécules d'ADN à deux brins est soumise à une dénaturation suivie d'une rénaturation, de manière à former un mélange des molécules d'ADN homo- et d'hétéroduplex. Afin de permettre une quantification des molécules d'AN détectées, il est prévu d'effectuer une étape de purification entre les étapes a) et b), ainsi que b) et c).

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