Thermostable enzyme promoting the fidelity of thermostable DNA polymerases-for improvement of nucleic acid synthesis and amplification in vitro
AssigneesRoche Diagnostics GmbH, Roche Molecular Systems, Inc.
InventorsWaltraud Ankenbauer, Frank Laue, Harald Sobek, Michael Greif
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Abstract
A purified thermostable enzyme is derived from the thermophilic archaebacterium Archaeoglobus fulgidus. The enzyme can be native or recombinant, is stable under PCR conditions and exhibits double strand specific exonuclease activity. It is a 3′–5′ exonuclease and cleaves to produce 5′-mononucleotides. Thermostable exonucleases are useful in many recombinant DNA techniques, in combination with a thermostable DNA polymerase like Tag especially for nucleic acid amplification by the polymerase chain reaction (PCR).
Claims4
A composition comprising a first thermostable Archeoglobus fulgidus enzyme exhibiting 3′–5′ exonuclease activity but essentially no DNA polymerase activity and a second enzyme exhibiting DNA polymerase activity, wherein said first thermostable Archeoglobus fulgidus enzyme is active under the incubation and temperature conditions of a polymerase chain reaction and is encoded by DNA which hybridizes to a primer having a sequence as shown in SEQ ID NO:
- 1 and a primer having a sequence as shown in SEQ ID NO:
- 2, said composition enhancing the fidelity of an amplification process in comparison to the use of the single second enzyme.
Dependencies are inferred from the claim text.