Qiagen Gmbh

Qiagen Manchester Limited

Qiagen Hamburg Gmbh

Holger Engel

Christian Korfhage

Ralf Peist

Dirk Löffert

Dirk Loeffert

Francesca Di Pasquale

Subrahmanyam Yerramilli

Martin Kreutz

Martin Lee

Jordi Carrera Fabra

Ralf Himmelreich

Anna Comengés Casas

Thomas Rothmann

Andreas Missel

Nicola Jo Thelwell

Andy Wende

Alexander Azzawi

Sascha Strauss

Rainer Dahlke

Sarah Fakih

Corinna Küppers

Janina Cramer

Thomas Laue

Diane Rachel Lee

Ricard Martín Blanco

Thomas Rohthmann

A microfluidic system is presented that includes a cartridge, a container, and a lid for the container. The cartridge includes a plurality of microfluidic channels coupled to one or more chambers. The container holds dry chemicals and includes a housing with a first opening and a second opening smaller than the first opening. The container is designed to be inserted into an opening of the cartridge, such that the container is independently secured within the opening. The insertion of the container allows for the container to be fluidically coupled with a microfluidic channel of the plurality of microfluidic channels via the second opening. The lid includes a column that extends from the lid into the container.

Dry chemistry container

The method for isolating nucleic acids from a food sample comprising the following steps: a) obtaining a food enrichment culture; b) transferring a portion of the food enrichment culture into a reaction vessel thereby providing a food enrichment sample and providing a water-immiscible phase in contact with the food enrichment culture; c) lysing the food enrichment sample to provide a lysed sample; d) isolating nucleic acids from the lysed sample. Food enrichment culture samples are known to contain high concentrations of organic and/or liposoluble inhibitors. By contacting the enrichment culture sample with a water-immiscible phase before the actual DNA extraction procedure starts, part of the lipophilic inhibitors is expected to cross the phase interface due to an enhanced solubility in the organic phase and thereby become depleted. The water-immiscible phase provided according to the invention interacts directly with the sample material throughout bacteria lysis and DNA extraction thereby optimizing the subsequent DNA purification processes due to the depletion of food-borne inhibitors. This method yields nucleic acids, in particular DNA that is of an improved quality and purity to be used in a subsequent PCR reaction to detect pathogen nucleic acids in the isolated nucleic acids.

Method for isolating nucleic acids from a food sample

This invention relates to a process for synthesis of a cDNA in a sample, in an enzymatic reaction, whereby the process comprises the steps: simultaneous preparation of a first enzyme with polyadenylation activity, a second enzyme with reverse transcriptase activity, a buffer, at least one ribonucleotide, at least one deoxyribonucleotide, an anchor oligonucleotide; addition of a sample that comprises a ribonucleic acid; and incubation of the agents of the previous steps in one or more temperature steps, which are selected such that the first enzyme and the second enzyme show activity. The invention further relates to a reaction mixture that comprises a first enzyme with polyadenylation activity, a second enzyme with reverse transcriptase activity, optionally a buffer, optionally at least one ribonucleotide, optionally at least one deoxyribonucleotide, and optionally an anchor oligonucleotide. Moreover, the invention relates to a kit that comprises a corresponding reaction mixture.

Reaction mixtures for forming cDNA from an RNA template

The present invention relates to a method for quantifying and/or detecting one or more nucleic acids of a genome in a sample, wherein in an amplification reaction, (i) a first nucleic acid is amplified, the locus that is amplified is a multicopy locus (MCL) within the genome, wherein the locus shares at least 80% sequence identity to a sequence according to SEQ ID NO. 1 over a stretch of 80 base pairs, and wherein the multicopy locus has copies on at least two different chromosomes, (ii) a second nucleic acid that has been added as an internal control (IC) is also amplified, and (iii) the amount of amplification product from the amplification of the first nucleic acid is determined.

Method for quantifying human DNA using an internal control

The present invention concerns a method for a polymerase chain reaction, in which a template nucleic acid, at least one primer, deoxyribonucleoside triphosphates as well as a DNA polymerase with proofreading activity are used. In addition, according to this invention, at least one target substrate is added to the polymerase chain reaction, whereby the efficiency of the DNA polymerase with proofreading activity is significantly increased. Any molecule that reduces or, in the optimal case, blocks the 3',5&#x2032;-exonuclease activity of the DNA polymerase used is suitable as target substrate. Technical solutions for the added substrate (target substrate) are in particular single stranded, linear oligonucleotides, hairpin oligonucleotides and RNA and DNA molecules. Furthermore, a kit is disclosed which comprise the required reagents for the implementation of the method according to the invention.

Method for polymerase chain reactions with use of a DNA polymerase with proofreading properties

The invention provides for a method for quantifying one or more nucleic acids of a genome in a sample comprising the steps of, (a) amplifying a first nucleic acid to be quantified, (b) determining the amount of said first nucleic acid by comparison of the amount of amplification product from said first nucleic acid with at least one amplification product from a second template nucleic acid, (c) wherein said second template nucleic acid was generated using whole genome amplification and wherein the starting concentration of the second template nucleic acid is known.

Method for quantifying human DNA

This invention relates to a process for synthesis of a cDNA in a sample, in an enzymatic reaction, wherein the process comprises the steps: simultaneous preparation of a first enzyme with polyadenylation activity, a second enzyme with reverse transcriptase activity, a buffer, at least one ribonucleotide, at least one deoxyribonucleotide, an anchor oligonucleotide; addition of a sample that comprises a ribonucleic acid; and incubation of the agents of the previous steps in one or more temperature steps, which are selected such that the first enzyme and the second enzyme show activity. The invention further relates to a reaction mixture that comprises a first enzyme with polyadenylation activity, a second enzyme with reverse transcriptase activity, optionally a buffer, optionally at least one ribonucleotide, optionally at least one deoxyribonucleotide, and optionally an anchor oligonucleotide. Moreover, the invention relates to a kit that comprises a corresponding reaction mixture.

Method of polyadenylation and CDNA synthesis in a single reaction

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 present invention relates to a method for amplifying and detecting nucleic acid sequences in a reaction cartridge comprising, (i) providing a sample comprising at least one nucleic acid molecule, (ii) in a first reaction chamber of the cartridge providing reagents for an amplification reaction, (iii) mixing the sample with the amplification reagents, (iv) amplifying the at least one nucleic acid in the first reaction chamber of the cartridge, (v) transferring at least parts of the amplification reaction into a second and third reaction chamber of the cartridge each comprising a probe set and performing a melting point analysis in order to determine which of the probes has specifically bound a nucleic acid.

Detection of multiple nucleic acid sequences in a reaction cartridge

The present invention is related to normalized quantification of nucleic acids and to the normalization of quantities of nucleic acids in samples, e.g. mixtures of nucleic acids. The present invention relates to method for the normalization of the quantity of a nucleic acid to be quantified in a sample to the total quantity of nucleic acid in the sample; or to the total quantity of a specific class of nucleic acid in the sample.

Method of normalized quantification of nucleic acids using anchor oligonucleotides and adapter oligonucleotides

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 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.

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Hilden, Germany

Holger Engel