Roche Diagnostics Operations, Inc.

Boehringer Mannheim Gmbh

Roche Molecular Systems, Inc.

Roche Diagnostics Gmbh

F. Hoffmann-la Roche Ag

Christian Birkner

Herbert Von Der Eltz

Klaus Muehlegger

Axel Heuser

Armin Reidel

Herbert Waldman

Frank Bergmann

Joachim Siedel

Christine Markert-Hahn

Monika Soukupova

Hans-Joachim Hoeltke

Michael Harold Town

The present disclosure provides a method and system for the PCR amplification of a target sequence which suppresses non-specific amplification products. The disclosure concerns the use of a primer pair optimized to amplify a nucleic acid of a contaminant in the background of genomic DNA of a first organism. When DNA from a second organism suspected for comprising the contaminant is subjected to the same PCR-based amplification reaction, detection sensitivity and specificity of the contaminant is enhanced when an amount of genomic DNA of the first organism is present in the amplification reaction.

Detection of bacterial (Mollicutes) contamination

The invention provides a composition containing an aqueous liquid phase, bacterial cells, and an ionic compound dissolved in the liquid phase. The ionic compound is selected from the group consisting of 1-butyl-3-methyl-imidazolium-thiocyanate, 1-butyl-3-methyl-imidazolium-2(2-methoxy-ethoxy)ethylsulfate, 1-methyl-1-[4-(3-methyl-3H-imidazol-1-ium)-but-1-yl]-3H-imidazolium-di(toluylsulfate), and 1-butyl-3-methyl-imidazolium-octylsulfate. The compositions of the invention are advantageously used for preparing lysates of biological cells, particularly bacterial cells.

Reagents for lysis of bacterial cells

Disclosed are water-soluble ionic liquids suitable for promoting adsorption of nucleic acids to a solid phase. The use thereof, particularly methods for the isolation of nucleic acids from an aqueous solution, as well as kits for performing those methods are disclosed.

Isolation and purification of nucleic acids with a solid phase

Disclosed is a method for analyzing nucleic acids in a sample comprising the following steps: (i) adding a DNA binding dye containing a benzothiazolium or benzoxazolium group to the sample to be analyzed, (ii) carrying out a polymerase chain reaction, (iii) applying the sample to a gel matrix, (iv) separating the nucleic acid molecules according to their size by applying a voltage and (v) excitation with light of a suitable wavelength for the optical visualization of the nucleic acid/DNA binding dye complexes.

Detection of PCR products in gel electrophoresis

Biscationic organic compounds are disclosed which promote adsorption of nucleic acids from an aqueous solution to a solid phase such as silica. Adsorption takes place under low salt conditions. Further disclosed are methods and kits suitable for nucleic acid isolation from aqueous solutions.

Nucleic acid adsorption under low-salt conditions

The present invention is directed to the use of a benzimidazolium compound comprising a side chain at at least one of its N-residues, said chain being either a Cn-alkyl or a substituted Cn-alkyl, characterized in that preferably n is &#x2267;3, as an additive for a nucleic acid amplification reaction.

Benzimidazolium compounds and salts of benzimidazolium compounds for nucleic acid amplification

The invention is related to the detection of a methylated cytosine in a nucleic acid wherein guanidinium hydrogen sulfite is used for the preparation of a solution containing guanidinium ions and sulfite ions and subsequent modification of the nucleic acid. Thereby, a non-methylated cytosine is converted to uracil. The invention further discloses kits for performing the methods of the invention.

Method for bisulfite treatment

Nucleotides labelled with an infrared dye and their use in nucleic acid detection

Blood substitute suppression by peroxides

Process for the production of oligonucleotides of formula II, in which the exocyclic amino groups of the bases adenine, guanine, cytosine, 7-deazaadenine and 7-deazaguanine carry N-phenylacetyl groups, are used for oligonucleotide synthesis, wherein in a first step a starting nucleotide is bound to a solid carrier, subsequently the desired oligonucleotide is synthesized by stepwise coupling with appropriately activated further monomeric nucleotide building blocks of the general formula I with the above-mentioned meanings, if desired, trivalent phosphorus is oxidized to pentavalent phosphorus during and after the synthesis, the oligonucleotide is cleaved from the carrier and the 5' protecting groups are cleaved off. The phenylacetyl functional groups that protect exocyclic NH.sub.2 groups of the bases can be cleaved off in a mild way with penicillin amidohydrolase (EC 3.5.1.11).

Growing community of inventors

Uffing, Germany

Christian Birkner