Abstract: Provided is a method for analysing the expression of one or more biomarker RNA molecules, comprising (A) isolating RNA from circulating tumor cells obtained from a subject, determining the expression of at least one biomarker RNA molecule in the isolated RNA and providing an expression profile based on the results; (B) isolating RNA from extracellular vesicles obtained from the subject, determining the expression of at least one biomarker RNA molecule in the isolated RNA and providing an expression profile based on the results; and (C) using the expression profiles determined in (A) and determined in (B) for a combined analysis of the results. Such combined analysis of the CTC and EV expression profiles enhances the prognostic and predictive value of the obtained results and can provide valuable diagnostic, prognostic and/or predictive information. The present method can thus be used as improved diagnostic, prognostic and/or predictive aid in the management of cancer patients.

Abstract: A kit for use in assessing the status of nucleic acid degradation and/or the integrity of one or more nucleic acids in a sample by amplifying at least two overlapping regions within at least one locus and detecting the amount of the at least two amplification products. The kit includes a primer and at least two probes that bind under stringent conditions to a sequence that shares at least 80% sequence identity to a sequence selected from the group of sequences consisting of SEQ ID NO. 6 to SEQ ID NO. 47 over a stretch of 80 base pairs, or to a reverse complement thereof. One of the at least two probes binds to one of the at least two overlapping regions and the other of the at least two probes binds to a non-overlapping region.

Abstract: A phenol-free method for isolating a nucleic acid from a sample is provided, said method comprising the following steps: a) adding a precipitation buffer to a sample to prepare an acidic precipitation mixture wherein said precipitation buffer comprises a metal cation precipitant and a buffering agent, has a pH value of 4.0 or less and does not comprise an organic solvent selected from aprotic polar solvents and protic solvents and wherein the acidic precipitation mixture comprises the metal cation precipitant in a concentration of less than 200 mM and precipitating proteins; b) separating the precipitate from the supernatant, wherein the supernatant comprises small RNA having a length of less than 200 nt and large RNA having a length of at least 1000 nt; and c) isolating a nucleic acid from the supernatant. The present method allows to avoid the use of organic solvents during protein precipitation. Also provided is a precipitation buffer.

Abstract: The present invention pertains to a method for isolating a target nucleic acid including small target nucleic acids from a sample, said method comprising at least the following steps a) binding at least a portion of the target nucleic acid including small target nucleic acids to a nucleic acid binding solid phase comprised in a column by passing the sample through said column, b) performing an enzymatic and/or chemical treatment on the nucleic acid binding solid phase while the target nucleic acid is bound to said solid phase, c) collecting at least a portion of the small target nucleic acids released from the solid phase during said treatment of step b) as flow-through, d) contacting said flow-through which comprises small target nucleic acids mixed with a recovery solution with a nucleic acid binding solid phase for binding the contained small target nucleic acids to said nucleic acid binding solid phase, e) optionally performing an elution.

Abstract: The present invention provides methods and composition suitable for stabilizing cell-containing samples such as blood samples. The stabilizers used are primary or secondary carboxylic acid amides.

Abstract: The invention is in the field of regulation of enzymatic activity in nucleic acid modifying reactions. It describes a method of regulating enzymatic activity by adding chelating agents to the reaction composition and exploits the fact that both the binding of divalent cations to these chelating agents and the pH of commonly used buffers is temperature dependent. PCR experiments that are hampered by non-specific side products can be regulated such that the target sequence is amplified in a more specific manner.

Abstract: The invention relates to a method of neighbor influence compensation between a plurality of objects in at least one digital image, wherein the at least one digital image contains image information about a plurality of objects. Each of the plurality of objects is configured to receive at least one molecule comprising genetic information, wherein the at least one molecule is configured to receive a fluorescent compound, and the at least one digital image is taken by an optical imaging system during emission of electromagnetic radiation of the fluorescent compounds received by the at least one molecules.

Abstract: Modular flow cells, devices with modular flow cells, and methods of sequencing using modular flow cells, as well as systems and kits including modular flow cells, are described, permitting sequencing wherein less than the full capacity for sequencing is desired.

Abstract: PCR that allows the researchers to amplify a desired DNA requiring only tiny amounts of sample. Such amplification reactions are technically challenging and are often hampered by several practical issues such as the presence of PCR inhibitors, sample degradation and low quantities of said sample. The invention addresses these issues with a method for evaluating the amplification efficiency and/or the presence of inhibitors and/or degradation and/or performing a quantification of a nucleic acid in a real-time amplification reaction comprising: optionally amplifying in a reaction composition a first target nucleic acid using a first primer pair in a real-time amplification reaction, (i) amplifying in said reaction composition one or more second internal nucleic acid control templates (IC) with a length of between 50 and 2000 nucleotides, wherein the second nucleic acid has a sequence selected from the group of: i) SEQ ID NO.

Abstract: The present invention relates to a chromatographic device for isolating and purifying nucleic acids, preferably genomic DNA, by gel filtration chromatography, a method for isolating and purifying nucleic acids, preferably genomic DNA, using this device and a kit comprising this device.

Abstract: The invention refers to a novel method of preparing strand-specific RNA-sequencing libraries that can be used to identify DNA coding and non-coding strands that are transcribed to RNA. Such strand-specific RNA-sequencing libraries are especially useful in discovering anti-sense RNA and non-coding RNA. Random primer oligonucleotides, covalently coupled to a moiety, which blocks ligation, are used for RT reaction or the subsequent generation of the second DNA strand so that only one strand of the generated double-stranded DNA is ligated to sequencing adapters at the 5? nucleotide and sequenced by paired-end sequencing.

Abstract: The present invention is directed to a method for immobilizing nucleic molecule on solid support and to a use of a nucleic acid non-immobilized primer in combination with a nucleic acid primer linked to a solid support in said a method.

Abstract: The present invention provides a method for preparing a sterilized composition suitable for stabilizing an extracellular nucleic acid population of a biological sample. Also provided are sterilizable compositions, wherein the compositions in sterilized form are suitable for stabilizing an extracellular nucleic acid population of a biological sample. Further useful methods, devices, kits and uses are also provided. Further sterilisable and sterilized compositions described herein are also suitable to stabilize intracellular nucleic acids (e.g. intracellular DNA such as genomic DNA and/or intracellular RNA) and cell characteristics, such as e.g. cell surface proteins and/or the cell morphology.

Abstract: Method and system for compensating intensity biases in a plurality of digital images. Each digital image of the plurality of digital images contains a plurality of objects and each of the plurality of objects is configured to receive at least one molecule comprising genetic information, wherein the at least one molecule is configured to receive one of at least a first fluorescent compound and a second fluorescent compound. A first digital image of the plurality of digital images is taken by an optical imaging system during emission of electromagnetic radiation by the first fluorescent compound, and a second digital image of the plurality of digital images is taken by the optical imaging system during emission of electromagnetic radiation by the second fluorescent compound.

Abstract: The invention relates to a method of neighbor influence compensation between a plurality of objects in at least one digital image, wherein the at least one digital image contains image information about a plurality of objects. Each of the plurality of objects is configured to receive at least one molecule comprising genetic information, wherein the at least one molecule is configured to receive a fluorescent compound, and the at least one digital image is taken by an optical imaging system during emission of electromagnetic radiation of the fluorescent compounds received by the at least one molecules.

Abstract: The present invention relates to a method for assessing the status of nucleic acid degradation and/or integrity of one or more nucleic acids in a sample, comprising the steps amplifying at least two overlapping regions within at least one locus (e.g. by heminested or nested PCR), and detecting the amount of the at least two amplification products through the use of at least two probes, wherein one probe binds to the region of overlap and the at least one other probe binds to a non-overlapping region. The invention further relates to a method of designing primers and/or probes for amplifying at least two overlapping regions within at least one locus, wherein the locus that is amplified is a single copy locus (SCL) or multicopy locus (MLC). The invention also relates to a primer and a primer pair for amplifying at least two overlapping regions from one nucleic acid template which is a multicopy locus present in 21 loci in the human genome. These primers and probes may be in a kit.

Abstract: The invention relates to nucleic acid probes, nucleic acid probe libraries, and kits for detecting, classifying, or quantitating components in a complex mixture of nucleic acids, such as a transcriptome, and methods of using the same. The invention also relates to methods of identifying nucleic acid probes useful in the probe libraries and to methods of identifying a means for detection of a given nucleic acid.