Polymerases for analyzing or typing polymorphic nucleic acid...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C12N 9/12 (2006.01) C12N 15/00 (2006.01) C12N 15/63 (2006.01) C12N 15/85 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2280001

The present invention provides methods for use in identifying, analyzing and typing polymorphic DNA fragments, particularly minisatellite, microsatellite or STR DNA fragments. In particular, the invention provides methods using DNA polymerases, more particularly thermostable DNA polymerases, and most particularly Thermotoga polymerases or mutants or derivatives thereof, whereby minisatellite, microsatellite or STR DNA molecules may be amplified and analyzed for polymorphisms. The invention also relatesto polymerases having reduced, substantially reduced or eliminated ability to add non-template 3' nucleotides to a synthesized nucleic acid molecule. In accordance with the invention, such reduction or elimination may be accomplished by modifying or mutating the desired polymerase.

La présente invention concerne des procédés d'identification, d'analyse et de typage de fragments d'ADN polymorphes, notamment des fragments minisatellites, microsatellites ou d'ADN STR. L'invention concerne plus particulièrement des procédés utilisant des ADN polymérases, plus spécifiquement des ADN polymérases thermostables et encore plus spécifiquement des polymérases Thermotoga, ou des mutants ou des dérivés de celles-ci. Les molécules minisatellites, microsatellites ou d'ADN STR peuvent être amplifiées et analysées en vue de détecter des polymorphismes. L'invention concerne également des polymérases ayant réduit, sensiblement réduit ou éliminé leur capacité à ajouter des nucléotides 3' non matriciels à une molécule d'acide nucléique synthétisée. Selon l'invention, une telle réduction ou élimination peut être effectuée par la modification ou la mutation de la polymérase souhaitée.

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