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AssigneesRoche Diagnostics GmbH, Roche Molecular Systems, Inc.
InventorsWaltraud Ankenbauer, Frank Laue, Harald Sobek, Michael Greif
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A purified thermostable enzyme is derived form 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 Taq especially for nucleic acid amplification by the polymerase chain reaction (PCR).
A method of preparing or amplifying DNA comprising incubating DNA with a composition comprising a first thermostable Archeoglobus fulgidus enzyme exhibiting 3′-exonuclease-activity but essentially no DNA polymerase activity and a second enzyme exhibiting DNA polymerase activity and lacking proofreading activity, wherein said first thermostable Archeoglobus fulgidus enzyme is active under the incubation and temperature conditions of a polymerase chain reaction, 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.