Methods and compositions for cloning nucleic acid molecules

Details Methods and compositions for cloning nucleic acid molecules Methods and compositions for cloning nucleic acid molecules

C - Chemistry, Metallurgy

12

P

C12P 19/34 (2006.01) C12N 15/10 (2006.01) C12N 15/66 (2006.01) C12N 15/70 (2006.01) C07K 16/40 (2006.01) C12N 9/99 (2006.01)

Patent

CA 2339791

The present invention is directed generally to methods facilitating the cloning of nucleic acid molecules. In particular, the invention relates to the use of polymerase inhibitors, including but not limited to anti- polymerase antibodies (such as anti-Taqantibodies) and fragments thereof, to inactivate residual polymerase activity remaining after the amplification (particularly via PCR) of a target nucleic acid molecule. The invention further provides compositions, particularly storage-stable compositions, comprising one or more components, such as one or more restriction endonucleases and one or more polymerase inhibitors, that are useful in cloning amplified or synthesized nucleic acid molecules by the above-described methods. The invention also relates to nucleic acid molecules produced by these methods, and to genetic constructs (such as vectors) and host cells comprising these nucleic acid molecules.

L'invention concerne des procédés permettant de clôner des molécules d'acide nucléique. En particulier, l'invention traite de l'utilisation d'inhibiteurs de la polymérase, qui comprennent, sans s'y limiter, des anticorps anti-polymérase (comme les anticorps anti-Taq par exemple) et des fragments de ces derniers pour inactiver l'activité résiduelle de la polymérase après l'amplification (en particulier par réaction en chaîne de la polymérase) d'une molécule d'acide nucléique cible. L'invention a aussi pour objet des compositions, en particulier, des compositions stables au stockage, comprenant un ou plusieurs composants, tels qu'une ou plusieurs endonucléases de restriction et un ou plusieurs inhibiteurs de polymérase qui présentent une grande utilité pour clôner des molécules d'acide nucléique amplifiées ou synthétisées selon les procédés susmentionnés. L'invention concerne également des molécules d'acide nucléique produites selon ces procédés, et des produits de recombinaison génétiques (tels que des vecteurs) et des cellules hôtes comprenant ces molécules d'acide nucléique.

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