Methods for the reduction of stutter in microsatellite...

C - Chemistry – Metallurgy – 12 – P

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C12P 19/34 (2006.01) C07H 21/02 (2006.01) C07H 21/04 (2006.01) C07K 5/00 (2006.01) C08H 1/00 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2446443

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 greater than 50 %; 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 sorbitol effective to reduce stutter relative to the amount of stutter observed in the absence of sorbitol. The invention also provides compositions containing sorbitol, kits for amplifying microsatellites having a G+C content of greater than 50 %, and methods of using all of the foregoing.

La présente invention concerne une méthode permettant de réduire les artefacts d'une réaction en chaîne de polymérase (PCR) dans l'amplification d'un microsatellite. Cette méthode consiste à fournir un échantillon renfermant un microsatellite doté d'un contenu G+C supérieur à 50 %, à mettre en contact ledit échantillon avec au moins une enzyme possédant une activité de polymérase d'acides nucléiques, et à faire incuber l'échantillon avec l'enzyme, pour une durée suffisante de temps, et dans des conditions permettant l'amplification du microsatellite. On réalise, alors, l'incubation en présence d'une quantité efficace de sorbitol afin de diminuer les artefacts de PCR afférents à la quantité d'artefacts de PCR constatés en l'absence de sorbitol. Ladite invention a également trait à des compositions contenant du sorbitol, à des kits d'amplification de microsatellites possédant un contenu G+C supérieur à 50 %, et à des méthodes d'utilisation correspondantes.

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