Producing copies of nucleic acids

C - Chemistry – Metallurgy – 12 – P

Patent

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C12P 19/34 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2191426

Amplification of nucleic acids using oligonucleotides immobilised on solid phase supports is disclosed. Target nucleic acid strands (in a sample to be analysed) are hybridised to the oligonucleotides and copy target strands (incorporating immobilised oligonucleotide) are produced using the target strands as templates. The target strands are denatured from the copy target strands and rehybridised to non-extended oligonucleotides. In the next step, new copy target (1) strands are synthesised from the oligonucleotides hybridised to the target strands, and simultaneously copy target (2) strands are synthesised using previously generated copy target (1) strands. Sequence of denaturing, rehybridising, and producing copy target (1) and copy target (2) strands is repeated as often as necessary to give the desired degree of amplification.

L'invention concerne l'amplification d'acides nucléiques à l'aide d'oligonucléotides immobilisés sur des supports en phase solide. On hybride des brins cibles d'acide nucléique (dans un échantillon à analyser) avec les oligonucléotides et on obtient des brins cibles de copie (comportant l'oligonucléotide immobilisé) à l'aide de brins cibles en tant que matrices. On dénature les brins cibles à partir de brins cibles de copie et on les hybride à nouveau avec des oligonucléotiques n'ayant pas subi d'extension. Dans l'étape suivante, on synthétise des nouveaux brins cibles (1) de copie à partir des oligonucléotides hybridés avec les brins cibles, et en même temps on synthétise les brins cibles (2) de copie à l'aide des brins cibles (1) de copie précédemment produits. On répète la séquence de dénaturation, de réhybridation et de production de brins cibles (1 et 2) de copie aussi souvent que nécessaire afin d'obtenir le degré souhaité d'amplification.

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