Amplicon melting analysis with saturation dyes

Details Amplicon melting analysis with saturation dyes Amplicon melting analysis with saturation dyes

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) C07D 279/16 (2006.01) C09B 23/04 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2501144

Methods are provided for nucleic acid analysis wherein a target nucleic acid that is at least partially double stranded is mixed with a dsDNA binding dye having a percent saturation of at least 50% to form a mixture. In one embodiment, the nucleic acid is amplified in the presence of the dsDNA binding dye, and in another embodiment a melting curve is generated for the target nucleic acid by measuring fluorescence from the dsDNA binding dye as the mixture is heated. Dyes for use in nucleic acid analysis and methods for making dyes are also provided.

L'invention concerne des procédés d'analyse d'acides nucléiques consistant à mélanger un acide nucléique cible au moins partiellement double brin avec un colorant se liant avec l'ADNds, et présentant un pourcentage de saturation d'au moins 50 % afin de former un mélange. Dans un mode de mise en oeuvre, l'acide nucléique est amplifié en présence du colorant se liant à l'ADNds, tandis que dans un mode de mise en oeuvre différent, une courbe de fusion est établie pour l'acide nucléique cible à partir de la mesure de la fluorescence émise par le colorant se liant à l'ADNds pendant le chauffage du mélange. L'invention concerne également des colorants convenant pour l'analyse des acides nucléiques et des procédés permettant de produire ces colorants.

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