Abstract

Microvesicles are enriched from a sample (for example exosomes) for subsequent isolation of biomolecules contained in the microvesicles, in particular RNA. A method involves: a) addition of an aqueous solution of salt of a polyuronic acid to the sample, b) addition of a substance which induces gel formation/pellet formation of the polyuronic acid, c) mixing of the sample and short incubation, d) centrifugation of the sample and removal of the supernatant, e) dissolving the pellet of gel piece, and 0 isolation of the biomolecules contained in the microvesicles, preferably RNA. Alginate is used as a preferred salt.

IPC Classes  ?

• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA

Abstract

The invention relates to magnetically separable polymer-based macro granules for carrying out immunoassays for detecting highly diverse analytes for medical, biological and biotechnological sectors. The size of the macroscopic granulate particles is between 0.5 mm and 10 mm in cross-section, preferably between 1 mm and 5 mm. Preferably, the macro granules can be magnetically separated. According to a preferred embodiment, the polymer-based macro granules are located in a pipette tip.

IPC Classes  ?

• G01N 33/543 - ImmunoassayBiospecific binding assayMaterials therefor with an insoluble carrier for immobilising immunochemicals

Abstract

The present application relates to a method and a test kit for carrying out a reaction for bisulphite modification of DNA in order to determine the patterns of methylation of the DNA, characterised in that, after a deamination reaction using bisulphite (e.g. sodium bisulphite or ammonium bisulphite) has taken place, the modified DNA is bound to a rough solid phase and all additional necessary method steps are carried out on this rough solid phase. Finally, the modified high-purity DNA is separated from this solid phase. The method can be carried out manually or automatically. The rough solid phase is a non-mineral material with a rough surface.

IPC Classes  ?

• C12Q 1/6806 - Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay

Abstract

The invention relates to magnetically separable polymer-based macro granules for carrying out immunoassays for detecting highly diverse analytes for medical, biological and biotechnological sectors. The size of the macroscopic granulate particles is between 0.5 mm and 10 mm in cross-section, preferably between 1 mm and 5 mm. Preferably, the macro granules can be magnetically separated. According to a preferred embodiment, the polymer-based macro granules are located in a pipette tip.

IPC Classes  ?

• G01N 33/543 - ImmunoassayBiospecific binding assayMaterials therefor with an insoluble carrier for immobilising immunochemicals

Abstract

The invention relates to the concentration and isolation of biological cells from a sample and/or the concentration of biological cells from a sample, followed by the isolation of nucleic acids from said cells, wherein the sample is brought into contact with a solid phase which has a rough or structured surface.

IPC Classes  ?

• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA

Abstract

The invention relates to a method for size-fractionated isolation of nucleic acids, characterized by the following steps: —a first binding buffer, which contains at least one chaotropic salt and at least one substance that raises the pH of the binding buffer, is added—in the absence of aliphatic alcohols—to a volume of the mixture of nucleic acids, —binding on a solid phase and separation of the nucleic acids bound by step a), —a second binding buffer, which has a lower pH than the binding buffer under Point a), or a nonionic surfactant or an alcohol or a mixture of nonionic surfactant and alcohol is mixed with the filtrate from step a), —binding on a solid phase and separation of the nucleic acids bound by step c), —washing and elution, according to known methods, of the nucleic acid isolated after steps a) and c), with the result that the nucleic acids isolated after step a) not only have a larger number of base pairs than the nucleic acids isolated under step c), but also that, both after both step a) and after step c), individual, particular nucleic acid fractions with a particular number of base pairs are isolated that were not isolated in the respective other step. The size ratios of the nucleic acid fractions can be controlled by changing the pH.

IPC Classes  ?

• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA
• C07H 1/08 - SeparationPurification from natural products
• C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids

Abstract

The invention relates to a method for enriching biomolecules and for removing the biomolecules from a biological sample, characterised in that, in the presence of particles, an alginate solution and salts of di- or polyvalent cations, or an acid are added to a biological sample, wherein an alginate-gel-biomolecule-complex is formed on the particles, which is removed via separation of the particle from the sample, and from which the biomolecules or the contents of the biomolecules are subsequently released. The biomolecules that are to be enriched include cell-free nucleic acids, viruses or subcellular microparticles. The method is an improved and simplified method compared to the method described in the patent specification DE 10 2008 023 297 B4.

IPC Classes  ?

• C07K 1/32 - ExtractionSeparationPurification by precipitation as complexes
• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA
• C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids

Abstract

Device and process for the automated extraction of nucleic acids, comprising a body which is able to be immersed partly or wholly into a reaction cavity, characterized in that at least the part immersed into the reaction cavity has a non-smooth surface. After lysis, a biological sample is admixed with an organic substance, preferably alcohols or ketones. This batch is then contacted with a material characterized by a non-smooth surface. In these circumstances, nucleic acids are adsorbed onto the surface of the material used. This is followed optionally by washing steps with known alcoholic washing solutions. After drying, the adsorbed nucleic acid is detached from the material by adding water or a low-salt buffer and can be used for downstream applications.

IPC Classes  ?

• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA
• G01N 1/40 - Concentrating samples
• B01L 3/02 - BurettesPipettes
• B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers

Abstract

The invention relates to a method for isolating nucleic acids from aqueous nucleic-acid-containing samples which comprise nucleic acids that are free or have been liberated by lysis. Before or after reduction in polarity of the aqueous solution, the sample is contacted with a solid phase having a rough or structured surface, and the nucleic acids precipitate on the solid phase and are subsequently removed from this aqueous solution with the solid phase. The rough or structured surface is preferably a non-smooth metal, plastic or rubber surface. The invention also provides a test kit and corresponding apparatus for isolating nucleic acids.

IPC Classes  ?

• C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA
• G01N 1/40 - Concentrating samples

Abstract

A device and method for extracting nucleic acids, comprising a hollow body, preferably a pipette tip, through which a fluid is guided, characterized in that a material with a rough or structured surface is arranged in this hollow body such that the material can be washed around with a fluid. After lysis of the sample and adjusting necessary binding conditions for the adsorption of the nucleic acids on the carrier material, the formulation is "pipetted past" the nucleic acid binding material located vertically in the pipette tip multiple times by means of a pipette process. The nucleic acids bind to the material. Subsequently, washing buffers are also "pipetted past" the nucleic acid binding material. A drying step subsequently takes place. Finally, the eluent is in turn "pipetted past" the nucleic acid binding material multiple times, said material being arranged vertically, and the bound nucleic acid is thereby dissolved. The nucleic acid is now available for necessary downstream application.

IPC Classes  ?

• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA
• G01N 1/40 - Concentrating samples

Abstract

The invention relates to a method for the size-fractionated isolation of nucleic acids, characterised by the following steps: - a first bonding buffering agent, containing at least one chaotropic salt and at least one substance which increases the pH value of the bonding buffering agent, is added to a volume of the mixture of nucleic acids (in the absence of aliphatic alcohols), - bonding to a solid phase and separation of the nucleic acids bonded by means of step a), - the filtrate from step a) is mixed with a second bonding buffering agent, which has a lower pH value than the bonding buffering agent in point a), or with a non-ionic surfactant, or with an alcohol, or with a mixture of a non-ionic surfactant and alcohol, - bonding to a solid phase and separation of the nucleic acids bonded by means of step c), - washing and elution of the nucleic acids isolated after steps a) and c) according to known methods, with the result that not only do the nucleic acids isolated after step a) have a greater number of base pairs than the nucleic acids isolated in step c), but that also individual, specific nucleic acid fractions with a specified number of base pairs are isolated, both after step a) and also after step c) which fractions are not isolated in the respective other step. The proportions of the nucleic acid fractions can be controlled by the alteration of the pH values.

IPC Classes  ?

• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA

Abstract

The invention relates to a simple method for enriching microvesicles from a sample (e.g. exosomes) in order to subsequently isolate the biomolecules, especially RNA, contained in the microvesicles and optionally detect same in a sensitive manner. The method comprises the following steps: a) adding an aqueous solution of a salt of a polyuronic acid to the sample; b) adding a substance which induces gel formation / pellet formation of the polyuronic acid; c) mixing the sample and brief incubation; d) subjecting the sample to centrifugation and removing the supernatant; e) dissolving the pellet or gel piece; f) isolating the ingredients, preferably RNA, biomolecules contained in the microvesicles in a known fashion. The preferred salt used is alginate.

IPC Classes  ?

• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA

Abstract

The invention relates to a method for detecting various analytes, characterized by the following steps: a) providing separation particles containing, on their surface, firstly means of binding the analyte to be identified and secondly means of separating the analyte bound to the particles; b) providing identification particles firstly having, on their surface, means for binding the analyte to be identified and secondly containing, on their surface or enclosed therein, means which are capable, after they have been detached or released from the particles, by virtue of their labeling, of generating a signal which serves for identification of the analyte; c) combining analyte, separation particles and identification particles; d) removing and washing the identification particles bound via the analyte by means of the separation particles; e) releasing the means which serve to identify the analyte, characterized in that the means which serve to identify the analyte are coupled reversibly to the identification particles and in that the identification molecules serve simultaneously for identification of the analyte and for detection.

IPC Classes  ?

• G01N 33/543 - ImmunoassayBiospecific binding assayMaterials therefor with an insoluble carrier for immobilising immunochemicals
• C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
• G01N 33/58 - Chemical analysis of biological material, e.g. blood, urineTesting involving biospecific ligand binding methodsImmunological testing involving labelled substances

Abstract

The invention relates to a device that allows a target nucleic acid to be detected in a homogeneous batch using two different detection formats. The device comprises at least two heatable sample blocks and a reaction cartridge which contains a base (1, 15) and at least one film (3, 4) sealing the base (1, 15), wherein the film (3, 4) comprises a surface (13, 14) that is not connected to the base (1, 15) and said surface (13, 14) forms a volume (16) for media transfer or at least one reaction chamber (10). The device can be used in particular for mobile gene diagnostics under field conditions.

IPC Classes  ?

• C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
• B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers
• B01L 7/00 - Heating or cooling apparatusHeat insulating devices

Abstract

The method for the real-time or end-point detection of nucleic acids is carried out using rehybridization probe systems. The novel probe system is characterized in that the system allows a homogenous and high-throughput nucleic acid detection that can be automated. Using the rehybridization probe system, a multiplex detection is also possible. In a particularly efficient embodiment of the invention, two probes are used for a qualitative or quantitative real-time measurement of specific target nucleic acids. In this manner, the detection sensitivity and signal strength can be substantially increased, and a target differentiation can be achieved.

IPC Classes  ?

• C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids

Abstract

The method and test kit are designed to qualitatively and quantitatively detect nucleic acid sequences in real time. The invention relates to the use of a DNA probe marked only with a fluorochrome (fluorophor), wherein the base marked with the fluorophor is always present in the vicinity of a guanine base and thus quenches the fluorophor without the use of a quenching dye. A mixture of duplexes is generated under hybridization conditions by means of primers, wherein the duplexes comprise the target nucleic acids deposited on the marked oligonucleotide (probe). By adding a polymerase having exonuclease activity, cutting the deposited marked oligonucleotide (probe) between the fluorophor marking and the guanine base, and thus terminating the quenching, a measurable fluorescence signal is generated.

IPC Classes  ?

• C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids

Abstract

The invention relates to the detection of specific nucleic acid sequences by means of fluorescence quenching. The basic principle of the detection is the steric exclusion of an antibody bond as the result of a nucleic acid target hybridizing to a nucleic acid probe to form a double strand. Once hybridization has taken place, the fluorophore of the probe is no longer accessible to a fluorescence-quenching antibody. The fluorescence which can be measured in the system is coupled directly to the presence, in the test sample, of a DNA sequence which is complementary to the probe.

IPC Classes  ?

• C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids

Abstract

The invention relates to a device, method and test kit for carrying out molecular-biological reactions, wherein the different components for the molecular-biological reactions are located on a solid carrier in different, spatially separated compartments prior to the start of the reaction. The carrier is preferably a porous filter disk made of polyethylene. Fields of application are the amplification of nucleic acids, for example PCR or RealTime PCR, the reverse transcription of RNA in DNA enzyme-substrate interactions or antigen-antibody interactions, or protein synthesis.

IPC Classes  ?

• C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids

Abstract

The invention relates to a method for concentrating bacteria, cells or viruses from biological samples, characterized in that the sample is converted into a liquid sample and treated with magnetic particles, and these magnetic particles together with the adsorbed bacteria, cells or viruses are subsequently removed from the liquid sample. The invention furthermore relates to a method for the subsequent isolation of nucleic acids from the concentrated bacteria, cells or viruses from biological samples, characterized in that the concentrated bacteria, cells or viruses are lysed by known methods and the nucleic acids are isolated by a known method.

IPC Classes  ?

• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA

Abstract

The invention relates to a method for providing constituents from biological samples, characterized in that liquid biological samples are treated with magnetic particles and these magnetic particles together with the bacteria, cells or viruses absorbed thereto are subsequently removed and the liquid sample, washed and subsequently lysed, and, after the lysis, the constituents of the bacteria, cells or viruses are present in the lysate.

IPC Classes  ?

• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA

Abstract

The present invention relates to a method and test kit for detecting specific nucleic acid sequences, comprising the steps of: 1. matrix-dependent new synthesis of the target nucleic acid; 2. target-specific probe hybridization; and 3. detection of the hybridization event. The invention is characterized in that, in the first step, an oligonucleotide 1, which is marked by a marker 1 and is entirely or partially complementary to the target sequence, acts as a primer in the matrix-dependent new synthesis of the target nucleic acid and, in the second step, an oligonucleotide 2, which is marked by a marker 2 and, owing to its melting temperature being lower than that of the oligonucleotide 1, is not involved in the first step, partially or completely hybridizes with the DNA new synthesis product of oligonucleotide 1. The detection of the hybridization reaction can take place both fluorometrically in the form of a homogeneous assay and, for verification of the result, subsequently immunologically. The detection reaction always takes place in time after the matrix-dependent new synthesis has been carried out.

IPC Classes  ?

• C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids

Abstract

The invention relates to a device that allows a target nucleic acid to be detected in a homogeneous batch using two different detection formats. The device comprises at least two heatable sample blocks and a reaction cartridge which contains a base (1, 15) and at least one film (3, 4) sealing the base (1, 15), wherein the film (3, 4) comprises a surface (13, 14) that is not connected to the base (1, 15) and said surface (13, 14) forms a volume (16) for media transfer or at least one reaction chamber (10). The device can be used in particular for mobile gene diagnostics under field conditions.

IPC Classes  ?

• B01L 7/00 - Heating or cooling apparatusHeat insulating devices
• B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers

Abstract

The object of the invention is a simple method for concentrating biomolecules (such as proteins or nucleic acids) from a sample for subsequent isolation of the biomolecules and alternatively sensitive detection. The method comprises the following steps: a) adding an aqueous solution of a salt of a polyuronic acid to the sample; b) adding a substance which induces gel formation / pellet formation of the polyuronic acid; c) mixing the sample and brief incubation; d) subjecting the sample to centrifugation and removing the supernatant; e) dissolving the pellet or gel piece; f) isolating the biomolecules in a known fashion. The preferred salt used is alginate.

IPC Classes  ?

• C07K 1/32 - ExtractionSeparationPurification by precipitation as complexes
• C08B 37/00 - Preparation of polysaccharides not provided for in groups Derivatives thereof
• C12N 7/00 - Viruses, e.g. bacteriophagesCompositions thereofPreparation or purification thereof
• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA

Abstract

Parallel isolation of a double-stranded nucleic acid and a single-stranded nucleic acid is possible from a sample that contains these acids, without separating the acids, by mixing the sample with a lysis buffer having high salt concentration or low salt concentration, or having a proteolytic enzyme. The sample that contains nucleic acid before its lysis, or the sample that has already been lysed or homogenized, is adjusted with a binding buffer in such a manner that the total nucleic acid is adsorbed onto a solid carrier. The binding buffer contains at least one non-ionic detergent in a high concentration. With the exception of the detergent, the sample contains no other non-acidic organic component miscible in water. The carrier with the adsorbed total nucleic acid is removed. The adsorbed total nucleic acid is washed and eluted.

IPC Classes  ?

• C12Q 1/70 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving virus or bacteriophage
• C12Q 1/6806 - Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay

Abstract

The invention relates to a mobile device system, comprising a hand-held device and a test kit for the mobile isolation of nucleic acids. The hand-held device comprises at least one sample block for inserting sample containers, a sample block holder with boreholes or recesses for accommodating the sample blocks, a device base with electronic control units for the sample blocks, a voltage source and a connection to the sample block holder, as well as a test kit for the isolation of nucleic acids. The hand-held device is characterized in that the sample blocks can be removed from the sample block holder and the sample block holder can be removed from the device base.

IPC Classes  ?

• B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers
• B01L 7/00 - Heating or cooling apparatusHeat insulating devices

Abstract

The present invention relates to a method and a test kit for the detection of specific nucleic acid sequences, with the steps of amplification, hybridization by means of probes, and the detection of the hybridization event, wherein the detection of the hybridization reaction takes place on a solid phase outside the reaction vessel of the amplification/hybridization.

IPC Classes  ?

• C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids

Abstract

The invention relates to a method and a test kit for the rapid detection of specific nucleic acid sequences, especially for the detection of mutations or SNPs (SNP = single nucleotide polymorphism), the detection reaction taking place in two steps. In a first step, the target-specific amplification reaction is carried out, together with the probe hybridising reaction using fluorescence-marked allele-specific amplification primers. In the second step, the fluorescence is detected by means of commercial fluorescence readers. The genotyping is carried out on the basis of the quotients of the end point fluorescence of the samples and the negative controls.

IPC Classes  ?

• C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids

Abstract

The invention relates to a method for parallel isolation of double- and single-stranded nucleic acids from samples containing said materials, without separating the double-and single-stranded nucleic acids. The samples are reacted with conventional lysis buffers (high salt concentrations, or low salt concentrations or with proteolytic enzymes). The sample containing nucleic acids before lysis thereof or after lysis or homogenisation is adjusted with an acidic binding buffer containing at least one non-ionic detergent in high concentration such that the total nucleic acids are adsorbed on a solid support.

IPC Classes  ?

• C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids

Abstract

The invention relates to a pharmaceutical composition that contains, as active components, at least one proteasome inhibitor and an inhibitor of protein folding enzymes. Said agents are suitable for treating acute and chronic infections having pathogenic viruses for humans and animals.

IPC Classes  ?

• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA

Abstract

The invention relates to a novel buffer formulation for the rapid separation, purification and highly-efficient recovery of long- and short-chain nucleic acids. In accordance with the invention, citric acid salts in combination with an alcohol are employed for this purpose.

IPC Classes  ?

• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA

Abstract

The invention relates to a universal and greatly simplified method of isolating nucleic acids from a wide range of starting materials comprising nucleic acids, in which method a combination of buffers with chaotropic components and buffers with nonchaotropic components is employed.

IPC Classes  ?

• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA

Abstract

The invention relates to a device and a method for automatically isolating nucleic acids from different nucleic acid-containing starting materials. The inventive reaction unit comprises a bottom part and a top part for pipetting liquids and is characterized in that the bottom part is composed of a reaction cavity with a permeable insertable filter grid while the top part represents a reaction cavity that can be fixed to the bottom part and is provided with a recess or a jacket for accommodating a magnet. The inventive method is suitable for the selective simultaneous isolation of genomic DNA of cellular RNA from any complex starting materials.

IPC Classes  ?

• C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids

Abstract

The invention relates to a universal and greatly simplified method as well as a formulation for isolating nucleic acids from different starting materials containing nucleic acids. Said formulation comprises at least one previously known buffer solution for proteolytically solubilizing biological samples, which is provided with no chaotropic or antichaotropic components, at least one alcoholic component and/or a detergent, a solid phase, and previously known detergent and elution buffers.

IPC Classes  ?

• C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA
• C07H 1/08 - SeparationPurification from natural products