Abstract: The present invention provides, inter alia, a method for generating a genome-wide epigenomic map, comprising a correlation between methylation variable CpG positions (MVP) and genomic DNA sample types. MVP are those CpG positions that show a variable quantitative level of methylation between sample types. Particular genomic regions of interest (ROI) provide preferred marker sequences that comprise multiple, and preferably proximate MVP, and that have novel utility for distinguishing sample types. The epigenic maps have broad utility, for example, in identifying sample types, or for distinguishing between and among sample types. In a preferred embodiment the epigenomic map is based on methylation variable regions (MVP) within the major histocompatibility complex (MHC), and has utility, for example, in identifying the cell or tissue source of a genomic DNA sample, or for distinguishing one or more particular cell or tissue types among other cell or tissue types.

Abstract: The present invention describes a method for the detection of nucleic acid sequences, which is characterized in that the following steps are conducted: a) at least one nucleic acid is bound to a solid phase; b) probe molecules are hybridized to the nucleic acids in a sequence-specific manner, whereby the probe molecules are provided with a cleavable bond and a mass label, which is specific for the probe molecule; c) removal of the unhybridized probe molecules; d) contacting of the hybridized probe molecules with a matrix, which cleaves said cleavable bonds and at the same time serves as the matrix in a MALDI mass spectrometer; e) detection of the mass label at those positions where the nucleic acid was bound.

Abstract: The present invention provides an improved method for the bisulfite conversion of DNA, and facilitates the analysis of cytosine methylation of genomic DNA. Novel combinations of denaturing solvents, new reaction conditions and new purification methods provide surprisingly efficacious methods for bisulfite conversion of DNA relative to prior art methods. The converted DNA may subsequently be analyzed by many different methods.

Abstract: Systems, methods and computer program products for guiding selection of a therapeutic treatment regimen or a preventive therapeutic treatment regimen are disclosed. The method comprises (A) providing to a computing device comprising a first knowledge base comprising information about a plurality of different methylation statuses at selected sites of the DNA in cells with a known disease or medical condition and/or healthy cells, a second knowledge base comprising a plurality of expert rules for evaluating and selecting a type of disease or medical condition based on the methylation status at selected sites of the DNA of a patient, (B) generating in said computing device a ranked listing of diseases or medical conditions based on the information about the methylation status at selected sites of the DNA of the patient, the first knowledge base and the second knowledge base.

Abstract: A method is disclosed providing analysis of the degree of methylation within nucleic acid samples, including the degree of methylation within CpG islands. After bisulfite treatment of a nucleic acid sample to convert cytosines to uracils, multiple species of paired oliogonucleotide primers and optionally a methylation insensitive reference primer pair are used to amplify target sequences within the sample by methylation specific PCR. Amplification of multiple primer pairs is combined with the use of a real time PCR. Amplificates of primer pairs are detected and quantified by comparison, thus allowing for a detailed, more specific, and quantifiable analysis of the methylation status within a complex CpG methylation pattern of a nucleic acid sample. Primer pairs and a kit are also disclosed.

Abstract: A method is described for controllably conducting complex PCR amplifications, wherein at least the following steps are conducted: a) PCR amplification with at least 50 primers of a first type (type 1) of different sequence, which are complementary to one of the strands of a random DNA sample, and also with a primer or a library of primers of a second type (type 2), which is complementary to the other strand of the DNA sample used, wherein the type 2 primers contain a first label (label 1); b) hybridizing of the amplified products to an oligomer array, which comprises oligonucleotides that hybridize to the primers utilized in the PCR reaction or to oligonucleotides that are complementary to these; or hybridizing of the amplified products to an oligomer array, which contains oligomers complementary to the primers utilized in the PCR reaction; c) length determination of the amplified products bound to the array by a second label (label 2) which can be correlated with the length of the respective DNA fragment,

Abstract: Chemically modified genomic sequences of genes associated with DNA replication, to oligonucleotides and/or PNA-oligomers for detecting the cytosine methylation state of genes associated with DNA replication which are directed against the sequence are disclosed. In addition, a method for ascertaining genetic and/or epigenetic parameters of genes associated with DNA replication is disclosed.

Abstract: The present invention relates to methods for detecting free floating nucleic acids, as present in not cellular bound nucleic acids in bodily fluids like plasma or serum fractions of human or animal blood or in any other tissue samples derived from the human or animal body in order to diagnose a cell proliferative disease. Specifically the invention relates to the detection of increased levels of nucleic acids in bodily fluids. Furthermore the invention allows to determine the source of the enriched DNA by measuring the ratio of DNA originating from a certain organ versus total DNA from other organs in a given bodily fluid sample by specifying the DNA's methylation pattern. This can be done with or without increasing the DNA concentration of a given biological sample. In a preferred embodiment a further analysis of this methylation pattern allows for the detection of the presence of tumourous or otherwise proliferative disease in said organ.

Abstract: Disclosed is a method and a set of oligonucleotides or PNA oligomers for detecting the methylation state of genomic DNA in a parallel manner. The DNA is treated with bisulphate and the thus chemically modified DNA is subsequently fragmented. In the next step, different fragments are amplified by means of synthetic primers. The amplificates are hybridized by means of oligonucleotides of a known sequence and are subsequently detected.

Abstract: An oligomer array with PNA (peptide nucleic acid) and/or DNA oligomers on a surface is described, which comprises oligomers of between 6 and 20 monomers or nucleobases each, whereby each of these contains at least one sequence of the general formula DDCGDD or of the general formula DDTGDD or of the general formula HHCGHH or of the general formula HHCAHH, wherein H indicates one of the bases: adenine (A), cytosine (C), or thymine (T) and D represents one of the bases: adenine (A), guanine (G) or thymine (T), and wherein the site of the oligomers on the surface is correlated with the sequence of the oligomers. The oligomer arrays according to the invention are used for the detection of cytosine methylations in genomic DNA.

Abstract: The present invention describes a method for the analysis of methylation patterns comprising the following steps: a) isolation of genomic nucleic acids from a biological sample, b) amplification of one or more target nucleic acids of said genomic nucleic acids in a manner whereby the methylation patterns of said genomic nucleic acids are maintained in the amplificate nucleic acid, c) performing mass spectrometry on said amplified nucleic acid or fragments thereof to obtain mass spectra; d) evaluating the obtained mass spectra and e) determining the methylation pattern and/or methylation status of the sample. The disclosed invention provides novel methods for the analysis of cytosine methylation patterns within genomic DNA samples. Said method comprises a methylation retaining enzymatic amplification of a test nucleic acid sample, followed by mass spectrometric analysis of the amplificate nucleic acids.

Abstract: The invention outlines a method for the methylation pattern retaining amplification of nucleic acid molecules. Furthermore the invention describes several devices for use in the methylation pattern retaining amplification of nucleic acid molecules.

Abstract: A sample holder is described, which is loaded with metal or glass slides. This arrangement serves for the analysis of DNA arrays in the MALDI-TOF mass spectrometer.

Abstract: A method is described for the detection of cytosine methylation in DNA samples, which permits the analysis of DNA to be investigated in the presence of large quantities of background DNA of the same individual. In the first step, a genomic DNA is chemically treated, preferably with a bisulfite (=disulfite, hydrogen sulfite), in such a way that cytosine is converted into a base that is different in its base pairing behavior in the DNA duplex, while 5-methylcytosine remains unchanged. Then segments of the sample DNA are amplified by means of a polymerase reaction. The amplificates are cleaved selectively by enzymes at those position which have a methylation state in the DNA sample that is not characteristic for the DNA to be investigated further, but which is characteristic for background DNA. The DNA that is not cleaved by enzymes is now amplified in another polymerase reaction, and in this way, the DNA to be investigated is concentrated relative to the background DNA that is present.

Abstract: A method for the detection of cytosine methylation in DNA samples is described, which is comprised of the following steps: First, a genomic DNA sample, which comprises the DNA to be investigated and background DNA, is chemically treated such that all unmethylated cytosine bases are converted to uracil, while the 5-methylcytosine bases remain unchanged. Then the chemically treated DNA sample is amplified with the use of at least 2 primer oligonucleotides as well as a polymerase, whereby the DNA to be investigated is preferred over the background DNA as the template, and in the last step, the amplificates are analyzed and the methylation status in the DNA to be investigated will be concluded from the presence of an amplificate and/or from the analysis of additional positions.

Abstract: The present invention describes a set of oligomer probes (oligonucleotides and/or PNA oligomers), which serve for the detection of the cytosine methylation state in nucleic acids. These probes are particularly suitable for the diagnosis of existing diseases by analysis of a set of genetic and/or epigenetic parameters.

Abstract: A method for the detection of cytosine methylation in DNA samples is described. First, DNA is extracted from a sample and bound to a surface. In the second step, a genomic DNA sample is preferably treated with a bisulfite (=disulfite, hydrogen sulfite), such that all unmethylated cytosine bases are converted to uracil, while the 5-methylcytosine bases remain unchanged. In the third step of the method, one or more oligonucleotides is (are) hybridized to the treated DNA as primers. In the fourth step of the method, the hybridized primer(s) is or are elongated in a polymerase reaction. Here, labeled guanine nucleotides are preferably utilized which are essentially incorporated only if cytosine bases were still present in the treated DNA. Consequently, the extent of incorporation of guanine bases and thus also the number of incorporated labels is proportional to the methylation in the DNA sample under investigation.

Abstract: A method for epigenetic knowledge generation which designs and synthesizes the chemical and/or biological components that determine the epigenetic parameters to be selected and measured is described. The value of these epigenetic parameters is determined, the steps of this procedure repeated and finally the results are stored. The present invention relates to a method of epigenetic knowledge generation comprising the steps of: a. selecting epigenetic parameters of interest; b. designing the chemical and/or biological components of the epigenetic measurement system, wherein the chemical and/or biological components determine the epigenetic parameters to be measured; c. synthesizing the variable chemical and/or biological components; d. measuring the value of the epigenetic parameters using the chemical and/or biological components; e. storing the results obtained by measurement; f. defining a subset of epigenetic parameters of interest based on the measurements; g. repeating steps a-d.

Abstract: Systems, methods and computer program products for guiding selection of a therapeutic treatment regimen or a preventive therapeutic treatment regimen are disclosed. The method comprises (A) providing to a computing device comprising a first knowledge base comprising information about a plurality of different methylation statuses at selected sites of the DNA in cells with a known disease or medical condition and/or healthy cells, a second knowledge base comprising a plurality of expert rules for evaluating and selecting a type of disease or medical condition based on the methylation status at selected sites of the DNA of a patient, (B) generating in said computing device a ranked listing of diseases or medical conditions based on the information about the methylation status at selected sites of the DNA of the patient, the first knowledge base and the second knowledge base.

Abstract: A set of oligonucleotides or PNA oligomers and a method which is suitable for the detection of cytosine methylations and SNPs in genomic DNA samples is described.