Methods for the reduction of stutter in microsatellite...

C - Chemistry – Metallurgy – 12 – P

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C12P 19/34 (2006.01) C07H 21/04 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2446626

The invention provides a method for reducing stutter in the amplification of a microsatellite comprising the steps of providing a sample comprising a microsatellite having a G+C content of 50% or less; contacting the sample with at least one enzyme having nucleic acid polymerase activity; and incubating the sample with the enzyme for a sufficient amount of time and under conditions sufficient to amplify the microsatellite; wherein the incubation is performed in the presence of an amount of betaine, sorbitol or mixtures thereof, effective to reduce stutter relative to the amount of stutter observed in the absence of betaine and/or sorbitol. The invention also provides compositions containing betaine and/or sorbitol, kits for amplifying microsatellites having a G+C content of 50% or less, and methods of using all of the foregoing.

L'invention concerne un procédé permettant de réduire les saccades dans l'amplification d'un microsatellite consistant à fournir un échantillon comportant un microsatellite présentant un contenu G+C de 50 % ou inférieur, mettre en contact l'échantillon avec au moins une enzyme dotée d'une activité polymérase acide nucléique et laisser incuber l'échantillon avec l'enzyme pendant une période de temps suffisante et sous certaines conditions suffisantes pour amplifier le microsatellite. L'incubation est réalisée en présence d'une quantité de bétaine, de sorbitol ou de mélanges de ceux-ci, efficace pour réduire les saccades par rapport à la quantité de saccades observée en l'absence de bétaine et/ou de sorbitol. Cette invention concerne également des compositions contenant de la bétaine et/ou du sorbitol, des ensembles d'amplification de microsatellites présentant un contenu G+C de 50 % ou inférieur et des procédés d'utilisation de tous les éléments susmentionnés.

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