Qiagen Gmbh

Dirk Löffert

Christian Korfhage

Ralf Peist

Ralf Himmelreich

Andreas Missel

Friederike Wilmer

Claudia Fritz

Kathleen Rieske

Patrick Baumhof

Christoph Erbacher

Holger Engel

Jie Kang

Nan Fang

Dominic O'Neil

Holger Wedler

Subrahmanyam Yerramilli

Martin Kreutz

Katja Decker

Erika Wedler

The present invention concerns a method for activating a nucleic acid for a polymerase reaction with the steps: (a) Heating a nucleic acid to a temperature of 55&#xb0; C. to 80&#xb0; C., (b) cooling the nucleic acid to a temperature at which a polymerase shows no substantial decrease in activity, and (c) starting the polymerase reaction by the addition of a heat-labile polymerase to the nucleic acid.

Method for activating a nucleic acid for a polymerase reaction

The invention relates to a method for the quantification of one or more nucleic acids in a sample, for example: making a sample available which contains at least one nucleic acid to be quantified, adding an oligonucleotide probe, the oligonucleotide probe comprising a sequence which can specifically hybridize to the nucleic acid to be quantified or to a common sequence of the nucleic acids to be quantified, incubating the sample under conditions which allow the hybridization of the oligonucleotide probe to the nucleic acid(s) to be quantified, incubating the sample under conditions which allow the extension of hybridized probes, the nucleic acid(s) serving as a template in each case, removing the non-hybridized probes from the sample and quantifying the hybridized oligonucleotide probes to measure the quantity of the nucleic acid(s) to be quantified. The invention also relates to a kit for carrying out said method.

Quantification of nucleic acids

The present invention concerns a method for inserting one or more tag sequences into a nucleic acid characterized by the following steps: (a) preparation of a template nucleic acid; (b) hybridization of at least one anchor sequence of at least one anchor oligonucleotide with one sequence section of the template nucleic acid; and (c) synthesization of a new strand of nucleic acid, which is partially complementary to the template nucleic acid and which contains a sequence complementary to the non-hybridized portion of the anchor oligonucleotide, e.g. to at least one tag sequence, on its 3' end.

Insertion of sequence elements into nucleic acids

The invention generally provides a method for the sample preparation for a subsequent preparation, processing or analysis method of a sample containing an at least one species of nucleic acid and/or one species of protein, whereby the method comprises the following steps: A) providing a sample which contains at least one species of a nucleic acid and/or of a protein, B) contacting the sample with a fluid or solid composition to produce a fluid sample preparation, whereby the composition contains at least a nitrogenous compound, which is selected from the group consisting of a) polyamines, b) amino acids, and oligo- and polypeptides, c) nitrogenous heterocyclic compounds, including homo-oder heteropolymeres, which comprise these nitrogenous compounds, d) amines of the type RRNR, whereby R, Rand Rare chosen independently from one another from the group consisting of H, C-C-alkyl groups and aryl groups, whereby R, Rand Rare not H simultaneously, e) carbonxylic acid amides, f) inorganic ammonium salts, g) ammonium groups containing inner salt compounds, h) antibiotica binding nucleic acid, i) compounds which bind in the small cavity of the DNA, and mixtures of two or more of these compounds. The invention provides in particular a method for the lysis of a biological sample, and methods for the stabilization of biomolecules, a method for the decrease of inhibiting effects in a sample containing biomolecules and a differential masking method.

Method for the treatment of a sample containing biomolecules

Disclosed are methods for the enhancement of the reactivation of thermostable reversibly inactivated enzymes comprising reactivating at least one thermostable reversibly inactivated enzyme in the presence of one or more nitrogen containing compounds.

Enhancing reactivation of thermostable reversibly inactivated enzymes

Novel thermostable chimeric nucleic acid polymerases and methods for their generation and use are disclosed. It is shown that these chimeric nucleic acid polymerases, such as DNA polymerases, can be constructed using enzymatically active domains, isolated from different proteins or chemically synthesized. It is demonstrated that chimeric nucleic acid polymerases of the present invention possess the chemical and physical properties of their component domains (e.g., exonuclease activity, thermostability) and that the chimeric polymerases are thermostable.

Thermostable chimeric nucleic acid polymerases and uses thereof

The invention relates to a method for processing RNA, in particular, a RNA reaction method and kits for carrying out said RNA reaction method.

Reverse transcription and amplification of RNA with simultaneous degradation of DNA

The invention relates to composition or a kit containing an enzyme that is reversibly inhibited by means of a chemical modification and an enzyme which is reversibly inhibited using non-covalent binding, the use of a mixture of enzymes reversibly inhibited in such a manner for processing or multiplying polynucleotides, and a method for specifically amplifying DNA by simultaneously using both types of reversibly inhibited enzymes.

Mixture of reversibly inhibited enzymes

The invention relates to a thermostable polymerase based on; DNA molecules which code for one such polymerase; expression vectors; host cells; methods for producing one such polymerase and the use thereof for polymerising nucleic acid, especially in the polymerase chain reaction.

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The invention generally provides a method for the sample preparation for a subsequent preparation, processing or analysis method of a sample containing an at least one species of nucleic acid and/or one species of protein, whereby the method comprises the following steps: A) providing a sample which contains at least one species of a nucleic acid and/or of a protein, B) contacting the sample with a fluid or solid composition to produce a fluid sample preparation, whereby the composition contains at least a nitrogenous compound, which is selected from the group consisting of a) polyamines, b) amino acids, and oligo- and polypeptides, c) nitrogenous heterocyclic compounds, including homo- oder heteropolymers, which comprise these nitrogenous compounds, d) amines of the type RRNR, whereby R, Rand Rare chosen independently from one another from the group consisting of H, C-C-alkyl groups and aryl groups, whereby R, Rand Rare not H simultaneously, e) carboxylic acid amides, f) inorganic ammonium salts, g) ammonium groups containing inner salt compounds, h) antibiotica binding nucleic acid, i) compounds which bind in the small cavity of the DNA, and mixtures of two or more of these compounds. The invention provides in particular a method for the lysis of a biological sample, and methods for the stabilization of biomolecules, a method for the decrease of inhibiting effects in a sample containing biomolecules and a differential masking method.

The present invention relates generally to the field of nucleic acid chemistry. More specifically, it relates to a method for enhancing the performance of coamplification reactions, e.g., multiplex PCR reactions.

Enhanced coamplification of nucleic acids

The invention generally provides a method for the sample preparation for a subsequent preparation, processing or analysis method of a sample containing an at least one species of nucleic acid and/or one species of protein, whereby the method comprises the following steps: A) providing a sample which contains at least one species of a nucleic acid and/or of a protein, B) contacting the sample with a fluid or solid composition to produce a fluid sample preparation, whereby the composition contains at least a nitrogenous compound, which is selected from the group consisting of a) polyamines, b) amino acids, and oligo- and polypeptides, c) nitrogenous heterocyclic compounds, including homo-older heteropolymeres, which comprise these nitrogenous compounds, d) amines of the type RRNR, whereby R, Rand Rare chosen independently from one another from the group consisting of H, C-C-alkyl groups and aryl groups, whereby R, Rand Rare not H simultaneously, e) carboxylic acid amides, f) inorganic ammonium salts, g) ammonium groups containing inner salt compounds, h) antibiotica binding nucleic acid, i) compounds which bind in the small cavity of the DNA, and mixtures of two or more of these compounds. The invention provides In particular a method for the lysis of a biological sample, and methods for the stabilisation of biomolecules, a method for the decrease of inhibiting effects in a sample containing biomolecules and a differential masking method.

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Düsseldorf, Germany

Dirk Löffert