Denaturing multi ion polynucleotide chromatography for...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/70 (2006.01) C07H 21/00 (2006.01) C07H 21/02 (2006.01) C12M 1/40 (2006.01) C12N 15/10 (2006.01) C12P 19/34 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2298019

The present invention is directed to improved methods for detection of mutations in DNA using Denaturing Matched Ion Polynucleotide Chromatography (DMIPC). The invention includes the following aspects: analysis of PCR amplification products to identify factors that affect PCR replication fidelity; design of PCR primers; selection of an optimal temperature for performing DMIPC; selection of the mobile phase composition for gradient elution; methods for column preparation and maintenance; and methods for preparing polynucleotide samples prior to chromatographic analysis.

La présente invention concerne des procédés améliorés de détection des mutations de l'ADN, au moyen d'une chromatographie par dénaturation d'ions appariés de polynucléotide (DMIPC). L'invention recouvre les aspects suivants: analyse des produits d'amplification par réaction en chaîne de la polymérase (PCR) afin d'identifier les facteurs affectant la fidélité de la réplication par PCR; conception des amorceurs de PCR; sélection d'une température adaptée à la réalisation de la DMIPC; sélection de la composition en phase mobile pour une élution graduée; procédés de préparation et d'entretien des colonnes; et procédés de préparation d'échantillons polynucléotidiques avant toute analyse par chromatographie.

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