Roche Diagnostics Operations, Inc.

Roche Molecular Systems, Inc.

Roche Diagnostics Gmbh

Boehringer Mannheim Gmbh

F. Hoffmann-la Roche Ag

Frank Laue

Waltraud Ankenbauer

Dieter Heindl

Gudrun Schmitz-Agheguian

Christine Ebenbichler

Bernhard Angerer

Vitaly Svetlichny

Elizaveta Bonch-Osmolovskaya

Harald Sobek

Michael Greif

Astrid Reiser

Gudrun Schmitz

Karl O Stetter

Rudiger Schmitt

Ursula Markau

Renate Kolb

Eva Walter

Andreas Huber

The present invention is directed to a composition comprising a DNA Polymerase which is preferably thermostable, Deoxynucleotides, at least one primer oligonucleotide or a pair of amplification primers, and randomized 5-8 mer oligonucleotide, characterized in that said oligonucleotide comprises a modification with an organic hydrophobic moiety Such a composition is specifically useful for performing hot start PCR.

Nucleic acid amplification in the presence of modified randomers

The present invention is directed to a synthetic peptide having a length of not more than 30 amino acids comprising a divalent cation binding site. Such a peptide according to the present invention is part of a composition for nucleic acid amplification and provides for a so-called hot start effect.

PCR hot start by magnesium sequestration

A purified thermostable enzyme is derived form the thermophilic archaebacterium. The enzyme can be native or recombinant, retains approximately 90% of its activity after incubation for two hours at 95&#xb0; C. in the presence of stabilizing agents and possesses 3'-5&#x2032; proofreading exonuclease activity. Thermostable DNA polymerases are useful in many recombinant DNA techniques, especially nucleic acid amplification by the polymerase chain reaction (PCR).

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The present invention is directed to a novel chemical compound comprising the structure [Xx-(CH2)m-phosphate-Yy]n, characterized in that 3&#x2266;m&#x2266;6, 30&#x2266;n&#x2266;60, each x and y is independently from each other 0 or 1, each X and Y is independently from each other any photometrically measurable entity; provided that the terminal X can also be an &#x2014;OH group or a phosphate group, and further provided that the terminal Y can also be an &#x2014;OH group. Such a compound can be used as a suitable hot start additive for PCR based amplification of nucleic acids.

Polyanion for improved nucleic acid amplification

A purified thermostable enzyme is derived from the thermophilic archaebacteriumThe enzyme can be native or recombinant, retains approximately 90% of its activity after incubation for two hours at 95&#xb0; C. in the presence of stabilizing agents and possesses 3'-5&#x2032; proofreading exonuclease activity. Thermostable DNA polymerases are useful in many recombinant DNA techniques, especially nucleic acid amplification by the polymerase chain reaction (PCR).

A purified thermostable enzyme is derived form the thermophilic archaebacterium. The enzyme can be native or recombinant, is stable under PCR conditions and exhibits double strand specific exonuclease activity. It is a 3'-5&#x2032; exonuclease and cleaves to produce 5&#x2032;-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).

Thermostable enzyme promoting the fidelity of thermostable DNA polymerases&#x2014;for improvement of nucleic acid synthesis and amplification in vitro

The present invention is directed to a new composition for performing a nucleic acid amplification reaction comprising (i) a thermostable DNA-Polymerase, (ii) a thermostable 3'-5&#x2032; Exonuclease, and (iii) at least one primer for nucleic acid amplification with a modified 3&#x2032; terminal residue which is not elongated by said thermostable DNA-Polymerase as well as methods for performing a PCR reaction using this composition. Furthermore, the method is directed to kits comprising such a composition.

Composition and method for hot start nucleic acid amplification

A purified thermostable enzyme is derived from the thermophilic archaebacterium. The enzyme can be native or recombinant, is stable under PCR conditions and exhibits double strand specific exonuclease activity. It is a 3'&#x2013;5&#x2032; exonuclease and cleaves to produce 5&#x2032;-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).

Thermostable enzyme promoting the fidelity of thermostable DNA polymerases-for improvement of nucleic acid synthesis and amplification in vitro

A DNA polymerase from a thermophilic eubacterium is provided. The DNA polymerase shows magnesium ion dependent reverse transcriptase activity and 3&prime;-5&prime; exonuclease activity. The invention also includes recombinant plasmids and transformed host cells capable of producing the enzyme. The enzyme is classified into class EC 2.7.7.7., a DNA nucleotidyl transferase DNA-directed type.

Thermostable DNA polymerase from carboxydothermus hydrogenoformans

The present invention provides a process for the reduction of non-specific star activities in the case of the specific cleavage of desoxyribonucleic acids by incubation with a restriction endonuclease in an appropriate buffer, wherein to the incubation batch is added an antibiotic which binds to the DNA on or near the star sequences of the enzyme but not within the specific recognition sequence of the restriction endonuclease.

Process for the reduction of star activities

The present invention provides a restriction endonuclease, characterized by the palindromic recognition sequence: ##STR1## and the cleavage position defined by the arrows. The present invention also provides a process for obtaining this new restriction endonuclease.

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Paehl-Fischen, Germany

Frank Laue