Abstract: The present invention provides a method for lysing a fixed biological sample, wherein the fixed biological sample comprises crosslinks between nucleic acid molecules and protein molecules due to the fixation, said method comprising (a) lysing the fixed biological sample, wherein lysis involves digestion with a proteolytic enzyme; (b) heating the lysed sample to reverse crosslinks; (c) adding a proteolytic enzyme and performing a proteolytic digestion; optionally wherein one or more additional treatment steps are performed between step (b) and step (c). The provided nucleic acids are of high yield and quality and can be purified from the lysed sample.

Abstract: The present invention relates to a composition for disrupting tissue material, the composition comprising solid disrupting particles in combination with at least one enzyme for enzymatic lysis and at least one chaotropic agent, as well as to a method for disrupting tissues material by simultaneously applying mechanical grinding or milling disruption and enzymatic digestion.

Abstract: The present invention pertains inter alia to a method for isolating poly(A) nucleic acids having a single stranded poly(A) stretch from a nucleic acid containing sample comprising: (a) providing a hybridization composition comprising: i) a nucleic acid containing sample; ii) a hybridization solution comprising: aa. a sodium salt; bb. a quaternary ammonium salt; wherein the components of the hybridization solution can be added as single solution to the sample or may be added separately in any order to the sample; iii) a capture probe capable of hybridizing to the poly(A) stretch of the poly(A) nucleic acids; and incubating said hybridization composition under conditions to form nucleic acid-hybrids between the poly(A) nucleic acids and the capture probe; (b) separating the formed hybrids from the remaining sample. The method is in particular suitable for efficiently isolating poly(A) RNA from various samples while avoiding carry-over of unwanted non-poly(A) nucleic acids such as rRNA.

Abstract: Provided is a method for isolating nucleic acids from a plant sample comprising (a) preparing a lysed sample wherein preparing comprises (i) lysing a plant sample by mechanically disrupting the plant sample in a lysis solution which comprises at least one chaotropic agent and one or more solid disrupting particles, and (ii) optionally clearing the lysate; (b) contacting the lysed sample with at least one protein precipitating agent and at least one inhibitor removing agent and providing a mixture; (c) obtaining a liquid phase from the mixture; and (d) isolating nucleic acids from the liquid phase. Also provided is a kit for use in such method.

Abstract: The present invention provides a lysis method for releasing microbial nucleic acids from microorganisms comprised in a plant sample, comprising mechanically disrupting the plant sample in a liquid lysis composition using at least two types of solid disrupting particles, wherein (i) the first type is provided by one or more disrupting particles having a size of at least 1.5 mm and (ii) the second type is provided by a plurality of disrupting particles having a size of 1 mm or less. Also provides is a method for isolating nucleic acids including microbial nucleic acids from a plant sample and kits.

Abstract: The present invention relates to a method for selectively depleting animal nucleic acids from non-animal nucleic acids in a sample, which comprises animal cells and at least one further type of cells, selected from microbial cells and plant cells or a combination thereof, to a lysis solution A to be used in and to a kit to carry out said method as well as to the use of a particular silica membrane as both, a filtration matrix for separating essentially intact microbial and/or plant cells from lysed animal cells and an adsorption matrix for nucleic acids, in particular in a method according to the present invention.

Abstract: The present invention relates to a method for selectively depleting animal nucleic acids from non-animal nucleic acids in a sample, which comprises animal cells and at least one further type of cells, selected from microbial cells and plant cells or a combination thereof, to a lysis solution A to be used in and to a kit to carry out said method as well as to the use of a particular silica membrane as both, a filtration matrix for separating essentially intact microbial and/or plant cells from lysed animal cells and an adsorption matrix for nucleic acids, in particular in a method according to the present invention.

Abstract: The present invention relates to a composition for disrupting tissue material, the composition comprising solid disrupting particles in combination with at least one enzyme for enzymatic lysis and at least one chaotropic agent , as well as to a method for disrupting tissues material by simultaneously applying mechanical grinding or milling disruption and enzymatic digestion.

Abstract: The present invention provides a poly(alkylene oxide) polymer based size selective DNA isolation method for isolating DNA molecules having a size above a certain cut-off value from a DNA containing sample, comprising (a) preparing a binding mixture comprising the DNA containing sample, at least one poly(alkylene oxide) polymer and at least one divalent cation, wherein said binding mixture has a pH that lies in the range of 8 to 10 and binding precipitated DNA molecules to a solid phase having an unmodified silicon containing surface, thereby providing a solid phase having bound thereto DNA molecules having a size above the cut-off value, wherein under the used binding conditions DNA molecules having a size which is less than the cut-off value substantially do not bind to the solid phase; (b) separating the bound DNA molecules from the remaining sample; —optionally washing the bound DNA molecules; and —optionally eluting the bound DNA molecules from the solid phase.

Abstract: The present invention pertains inter alia to a method for isolating poly(A) nucleic acids having a single stranded poly(A) stretch from a nucleic acid containing sample comprising: (a) providing a hybridization composition comprising: i) a nucleic acid containing sample; ii) a hybridization solution comprising: aa. a sodium salt; bb. a quaternary ammonium salt; wherein the components of the hybridization solution can be added as single solution to the sample or may be added separately in any order to the sample; iii) a capture probe capable of hybridizing to the poly(A) stretch of the poly(A) nucleic acids; and incubating said hybridization composition under conditions to form nucleic acid-hybrids between the poly(A) nucleic acids and the capture probe; (b) separating the formed hybrids from the remaining sample. The method is in particular suitable for efficiently isolating poly(A) RNA from various samples while avoiding carry-over of unwanted non-poly(A) nucleic acids such as rRNA.

Abstract: Methods and materials for determining the presence of at least one nucleic acid in a sample are provided, said methods comprising (1) a purification step using sequence specific hybrid capture; (2) an amplification step; and (3) a detection step using two separate sequence-specific polynucleotide probes. Also provided are nucleic acids comprising SEQ ID NO: 1 to SEQ ID NO: 727 and nucleic acid probes and probe sets comprising the same.

Abstract: Disclosed are compositions and methods for random amplification of nucleic acid sequences of interest using random-G-deficient primers. Also disclosed are methods of randomly amplifying a target nucleic acid sequence using random G-deficient primers alone or in combination with random, partially random, or specific primers.

Abstract: Methods and materials for determining the presence of at least one nucleic acid in a sample are provided, said methods comprising (1) a purification step using sequence specific hybrid capture; (2) an amplification step; and (3) a detection step comprising contacting the target nucleic acid with a plurality of detectably labeled nucleic acid detection probes, wherein each (a) bears a different detectable label from the other detection probes, and/or (b) has a different melting temperature from probes bearing the same detectable label. Also disclosed are compositions and kits for use in such a method.

Abstract: The present invention provides a method of preparing a target RNA depleted composition from an initial RNA containing composition, comprising a) contacting the initial RNA containing composition with one or more groups of probe molecules, wherein a group of probe molecules has the following characteristics: i) the group comprises two or more different probe molecules having a length of 100 nt or less; ii) the probe molecules comprised in said group are complementary to a target region of a target RNA; iii) when hybridized to said target region, the two or more different probe molecules are located adjacent to each other in the formed double-stranded hybrid; and generating a double-stranded hybrid between the target RNA and the probe molecules; b) capturing the double-stranded hybrid by using a binding agent which binds the double-stranded hybrid, thereby forming a hybrid/binding agent complex; c) separating the hybrid/binding agent complexes from the composition, thereby providing a target RNA depleted compos

Abstract: The invention relates to a method for enriching one or more target sequences of a deoxyribonucleic acid (DNA) in a composition, comprising the steps of providing a composition comprising one or more deoxyribonucleic acid (DNA) molecules, hybridizing to said one or more DNA molecules, one or more target specific ribonucleic acid (RNA) hybridization probes, thereby forming one or more RNA/DNA hybrids, capturing the RNA/DNA hybrids with one or more antibodies being specific for such RNA/DNA hybrids, thereby forming one or more RNA/DNA/antibody hybrids, isolating the one or more RNA/DNA/antibody hybrids, amplifying the one or more DNA molecules of the one or more RNA/DNA/antibody hybrids if necessary, and, optionally, sequencing the one or more DNA molecules of the one or more RNA/DNA/antibody hybrids or the amplification product, wherein the sequencing is preferably done by means of next generation sequencing.

Abstract: Methods and materials for determining the presence of at least one nucleic acid in a sample are provided, said methods comprising (1) a purification step using sequence specific hybrid capture; (2) an amplification step; and (3) a detection step comprising contacting the target nucleic acid with a plurality of detectably labeled nucleic acid detection probes, wherein each (a) bears a different detectable label from the other detection probes, and/or (b) has a different melting temperature from probes bearing the same detectable label. Also disclosed are compositions and kits for use in such a method.

Abstract: Disclosed are compositions and methods for random amplification of nucleic acid sequences of interest using random-G-deficient primers. Also disclosed are methods of randomly amplifying a target nucleic acid sequence using random G-deficient primers alone or in combination with random, partially random, or specific primers.

Abstract: Methods and kits for determining load of an infectious agent in a sample are described, comprising performing at least one hybridization assay and calculating the load of the infectious agent in the sample from a detected nucleic acid. In particular, the methods and kits disclosed determine the load of human papillomavirus (HPV) in a sample.

Abstract: Methods of selectively and rapidly identifying target nucleic acid molecules in large volumes of collection media where the target is present in a low concentration are disclosed. The methods can be used to identify, isolate, purify, or enrich a nucleic acid molecule containing a specific target sequence from a sample of nucleic acid molecules that do not contain the specific target sequence. Once isolated, the nucleic acid molecule containing a specific target sequence may be amplified or used in a variety of detection assays.

Abstract: Compositions, methods, and kits are provided for determining the presence of a target nucleic acid in a sample using synthetic probes.