Method and device for detecting a mutation in a nucleic acid...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) B01L 3/00 (2006.01) G01N 27/447 (2006.01) B01L 7/00 (2006.01)

Patent

CA 2426509

The present invention refers to methods and devices for the detection of polymorphisms in a nucleic acid sample (e.g. blood, sperm, saliva, cells, ...). To enhance the efficiency and the reliability of the known methods (e.g. DGGE, SSCP and TGGE) the amplification process (e.g. PCR) preceding the actual detection step is performed in or on the polyacrylamide gel. Multiple gradients (of chemical denaturants, thermal denaturants and of porosity of the gel matrix) are used for the separation of DNA fragments, by zone electrophoresis on gel slabs or by capillary electrophoresis.

Cette invention se rapporte à des procédés et à des dispositifs permettant de détecter les polymorphismes dans un échantillon d'acide nucléique (par exemple un échantillon de sang, de sperme, de salive, de cellules, etc). Pour améliorer l'efficacité et la fiabilité des procédés connus (tels que DGGE, SSCP et TGGE), le processus d'amplification (par exemple PCR) précédant la phase de détection effective est effectué dans ou sur le gel de polyacrylamide. De multiples gradients (de dénaturants chimiques, de dénaturants thermiques et de porosité de la matrice de gel) sont utilisés pour la séparation des fragments d'ADN, par électrophorèse de zone sur plaques de gel ou par électrophorèse capillaire.

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