Abstract: An improved method of in situ hybridization which relies on an improved formulation of the in situ hybridization buffer is described. In at least some formulations the buffer are non-toxic. The combination of Locked Nucleic Acid (LNA) comprising ISH probes and the improved ISH buffer are useful for detection of small non-coding RNA as well as in the manufacturing of ISH kits directed to the detection of such small non-coding RNA. Further disclosed is a method of semi-quantitative ISH and demonstration of the semi-quantitative ISHs diagnostic potential.

Abstract: An improved method of in situ hybridization which relies on an improved formulation of the in situ hybridization buffer is described. In at least some formulations the buffer are non-toxic. The combination of Locked Nucleic Acid (LNA) comprising ISH probes and the improved ISH buffer are useful for detection of small non-coding RNA as well as in the manufacturing of ISH kits directed to the detection of such small non-coding RNA. Further disclosed is a method of semi-quantitative ISH and demonstration of the semi-quantitative ISHs diagnostic potential.

Abstract: An improved method of in situ hybridization which relies on an improved formulation of the in situ hybridization buffer is described. In at least some formulations the buffer are non-toxic. The combination of Locked Nucleic Acid (LNA) comprising ISH probes and the improved ISH buffer are useful for detection of small non-coding RNA as well as in the manufacturing of ISH kits directed to the detection of such small non-coding RNA. Further disclosed is a method of semi-quantitative ISH and demonstration of the semi-quantitative ISHs diagnostic potential.

Abstract: An improved method of in situ hybridization which relies on an improved formulation of the in situ hybridization buffer is described. In at least some formulations the buffer are non-toxic. The combination of Locked Nucleic Acid (LNA) comprising ISH probes and the improved ISH buffer are useful for detection of small non-coding RNA as well as in the manufacturing of ISH kits directed to the detection of such small non-coding RNA. Further disclosed is a method of semi-quantitative ISH and demonstration of the semi-quantitative ISHs diagnostic potential.

Abstract: A method for simultaneous release and detection of nucleic acids from complex biological samples is described. The invention relates to the combined use of lysis buffers containing strong chaotropic agents such as guanidine thiocyanate to facilitate cell lysis and release of cellular nucleic acids and to the use of a novel type of bicyclic nucleotide analogues, locked nucleic acid (LNA) to detect specific nucleic acids released during lysis by nucleic acid hybridisation. In particular methods are described for the covalent attachment of the catching LNA-oligo. Novel methods for sample preparation of e.g. polyadenylated mRNA species are also presented. The invention further addresses reagents for performing the methods as well as reagents and applications of the method.

Abstract: The present invention relates to a method of detecting variant nucleic acid whose nucleotide sequence differs from one another at a single (or more) position(s). The method uses a set of chimeric oligonucleotides containing DNA monomers and monomers of a novel class of DNA analogues, locked nucleic acid (LNA). LNA oligomers obey the Watson-Crick base-pairing rules and form duplexes that are significantly more stable than similar duplexes formed by DNA. The “allele-specific” LNA-containing oligonucleotides wherein the LNA nucleotide(s) are found at the 3′ position can be extended by means of enzymes only where the nucleotide(s), which is/are terminal in direction of extension, is/are complementary to the corresponding nucleotides of the nucleic acid (of the one allele) to be detected. Thus discrimination between alleles without subsequent differential hybridization with labelled oligonucleotides is possible.

Abstract: The invention relates to therapeutic applications of LNA-modified oligonucleotides. In particular, the invention provides methods for treatment of undesired cell growth as well as treatment of inflammatory related diseases and disorders. Preferably, administration of an LNA-modified oligonucleotide modulates expression of a targeted gene associated with the undesired cell growth or an inflammatory related disease or disorder.

Abstract: The present invention relates to a method of detecting variant nucleic acid whose nucleotide sequence differs from one another at a single (or more) position(s). The method uses a set of chimeric oligonucleotides containing DNA monomers and monomers of a novel class of DNA analogues, locked nucleic acid (LNA). LNA oligomers obey the Watson-Crick base-pairing rules and form duplexes that are significantly more stable than similar duplexes formed by DNA. The “allele-specific” LNA-containing oligonucleotides wherein the LNA nucleotide(s) are found at the 3′ position can be extended by means of enzymes only where the nucleotide(s), which is/are terminal in direction of extension, is/are complementary to the corresponding nucleotides of the nucleic acid (of the one allele) to be detected. Thus discrimination between alleles without subsequent differential hybridization with labelled oligonucleotides is possible.

Abstract: A method for simultaneous release and detection of nucleic acids from complex biological samples is described. The invention relates to the combined use of lysis buffers containing strong chaotropic agents such as guanidine thiocyanate to facilitate cell lysis and release of cellular nucleic acids and to the use of a novel type of bicyclic nucleotide analogues, locked nucleic acid (LNA) to detect specific nucleic acids released during lysis by nucleic acid hybridisation. In particular methods are described for the covalent attachment of the catching LNA-oligo. Novel methods for sample preparation of e.g. polyadenylated mRNA species are also presented. The invention further addresses reagents for performing the methods as well as reagents and applications of the method.