Qiagen Gmbh

Holger Wedler

Roland Fabis

Dirk Löffert

Nan Fang

Thorsten Singer

Andreas Missel

Dominic O'Neil

Katja Heitz

Jan Petzel

Erika Wedler

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 invention relates to a chromatographic device for isolating and/or purifying double-stranded nucleic acids, preferably double-stranded DNA, from a mixture of such nucleic acids with single-stranded nucleic acids, oligonucleotides, mononucleotides, salts and/or other such impurities. The invention also relates to a method for chromatographically isolating and/or purifying same, and to a kit for this purpose.

Method and device for isolating and purifying double-stranded nucleic acids

The present invention relates to a method for purifying a defined amount of nucleic acids from a nucleic acid-containing sample, which has at least the following steps: (a.) contacting the nucleic acid-containing sample with a defined amount of a nucleic acid binding phase with the following features: (i) the nucleic acid binding phase has nucleic acid binding ligands that have at least one protonatable group; (ii) the nucleic acid binding ligands are bound to a carrier; (iii) the nucleic acid binding phase has a surface with a low charge density, wherein the amount of nucleic acids in the sample exceeds the binding capacity of the amount of nucleic acid binding phase used; (b.) binding of the nucleic acids to the nucleic acid binding phase at a pH (binding pH) that is below the pKs value of at least one of the protonatable groups; (c.) elution of the nucleic acids at a pH that is above the binding pH, wherein a defined amount of nucleic acids is obtained. Furthermore, corresponding kits and nucleic acid binding phases, which can be used for the purification of nucleic acids, are disclosed.

Nucleic acid purification method

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Hilden, Germany

Holger Wedler