Qiagen Gmbh

Andreas Missel

Nan Fang

Dirk Löffert

Jie Kang

Christian Korfhage

Ralf Peist

Holger Engel

Holger Wedler

Dirk L&ouml;ffert

Katja Heitz

The present invention is directed to methods for the generation of nucleic acids from an RNA template and further nucleic acid replication. Specifically, the invention is directed to the generation and amplification of nucleic acids by reverse transcriptase-polymerase chain reaction (RT-PCR).

Methods for RT-PCR comprising an anionic polymer

The present invention relates to methods and uses for the detection or quantification of newly-synthesized double-stranded target nucleic acid molecules in a sample during quantitative real-time polymerase chain reaction (qPCR) amplification. According to the invention, an intercalating dye recognizing double-stranded DNA molecules with higher affinity than single-stranded DNA molecules and a fluorophore-labeled oligonucleotide-probe being sequence specific for a target nucleic acid molecule are simultaneously employed, thus enabling quantification a specific target and total amount of a mixed nucleic acid population, and enabling assessing the cause of suboptimal PCR performance.

Nucleic acid detection and quantification

The present invention is in the fields of molecular biology. The present invention is directed to novel compositions, methods and kits useful for the generation of nucleic acids from an RNA template and further nucleic acid replication. Specifically, the invention is directed to the generation and amplification of nucleic acids by reverse transcriptase-polymerase chain reaction.

Composition and methods for RT-PCR comprising an anionic polymer

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

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 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

Novel compositions and methods useful for the generation of nucleic acids from a ribonucleic acid template and further nucleic acid replication are disclosed. It is shown that the generation and amplification of nucleic acids by methods that utilize two or more different polymerases, such as reverse transcriptase-polymerase chain reaction (RT-PCR), are dramatically more sensitive and efficient in the presence of a homopolymeric nucleic acid. Homopolymeric nucleic acids have been found to reduce or negate the inhibitory effect reverse transcriptases have on DNA polymerase activity. It is demonstrated that this inhibition-relieving effect of homopolymeric nucleic acids is general in nature; independent of the chemical species of homopolymer used, or the chemical composition of the polymerization reaction mixture.

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Hilden, Germany

Andreas Missel