40 results about "DNA Methylation Site" patented technology

The present invention relates to a tag-modified bisulfite genomic sequencing (tBGS) method developed for simplified evaluation of DNA methylation sites. The method employs direct cycle sequencing of PCR products at kilobase scale, without conventional DNA fragment cloning. The method entails subjecting bisulfite-modified genomic DNA to a second-round PCR amplification employing GC-tagged primers. The invention also relates to a method for identifying a patient at risk for lung cancer using the tBGS technique disclosed.

The invention discloses a DNA methylation marker used for diagnosis, screening, and prediction of male colorectal cancer. The DNA methylation marker is obtained via methylation of a combination of thefirst groups, or the first group with the second group, or the third groups, (1), +22, +27, and +29; (2), -24,-18, +89, and +92; and (3) +136; of three CpG sites in the front and back sequence of gene RPS24 transcription start site. The sequence number is based on sequence number in SEQ ID NO.1. The 143th site transcription start site A is recorded as +1. The invention also discloses a probe, a method, and a kit used for detecting the DNA methylation marker, and a computer module used for prediction of male colorectal cancer risk using the data of the DNA methylation sites.

The invention relates to a method for cancer diagnosis and/or cancer risk hazard degree assessment in subjects. The method comprises the following steps: a) determining DNA methylation states of samples from the subjects and samples from a healthy control group on at least one DNA methylation site; b) comparing the DNA methylation states of the samples from the subjects at the DNA methylation site with the DNA methylation states of the samples from the healthy control group at the corresponding DNA methylation site; and if the methylation states of the samples from the subjects and the samples from the healthy control group have remarkable difference, showing that the subjects have cancers or have cancer risks. The invention further provides a kit for the method provided by the invention.

The invention belongs to the field of the genetic engineering, and particularly provides a group of DNA methylation biomarkers for evaluating cardiac muscle cell damage and an application thereof. TheDNA methylation biomarker comprises 30 DNA methylation sites of 5 DMRs corresponding to 5 cardiac muscle differentiation key factors; and the five cardiac muscle differentiation key factors are GATA4, TNNT2, TNNI3, MEF2A and NKX2-5. Such type of the provided differential DNA methylation biomarkers is successfully developed to subvert the traditional biomarkers using protein as major means, a newsituation is created for the prevention and treatment of the cardiac muscle damage, and a reference is provided for the development of other disease biomarkers. The DNA methylation biomarkers are capable of evaluating the damage level of the cardiac muscle affected by environmental factors, and evaluating and forecasting the cardiotoxicity of environmental chemistry, and are the biomarkers with the prospect.

The invention discloses a dimensionality reduction method of DNA methylation spectrum for gastric cancer. The dimensionality reduction method includes the steps: selecting statistically significant methylation sites by analysis, filtering of insignificant methylation sites, maintaining single-factor, multi-factor statistically significant DNA methylation sites as seed methylation sites, further using consistent cluster analysis, obtaining the number of molecular subtypes with stable clustering results, and finally obtaining several cluster-specific methylation sites by screening and dimensionality reduction. The dimensionality reduction method of DNA methylation spectrum for gastric cancer can effectively reduce the dimensionality of high-dimensional and redundant DNA methylation data, canselect key DNA methylation sites from hundreds of thousands of DNA methylation sites as biomarkers, and has great significance for genetic research and drug development of gastric cancer.

The invention provides a DNA methylation data detection method and a device thereof based on seed sequence information. The method comprises the steps of building an index database; obtaining sequencing data of a target sample, and according to the preset seed sequence length, dividing the sequencing data to obtain divided seed sequence information; based on the index database, determining comparison candidate position information of each divided seed sequence information; conducting system evaluation on each comparison candidate position information to obtain a system evaluation result, and according to the system evaluation result, obtaining a DNA methylation locus of the target sample. Time of comparison operation which consumes most time in data analysis is greatly shortened, on the basis of guaranteeing the integrity of a methylation detection area locus, the utilization rate of data is increased by a large margin, the operation efficiency and accuracy of the data are improved bya large margin, and great convenience is brought to further research on DNA nucleobase modification information for scientific research staff in the field of life science.

The invention discloses a colorectal cancer DNA methylation marker, a method for detecting the colorectal cancer by using the colorectal cancer DNA methylation marker, and a kit. The marker is methylation of gene FXYD1, DZIP3, RPS24, AKT2, UBTF, DAGLB, LPAR6, TP63, CX3CR1 and TACC2 sequences. The invention also discloses a probe, a method and a kit for detecting the DNA methylation marker, and a computer module for predicting the colorectal cancer occurrence risk by utilizing data of a DNA methylation site.

The invention provides a method for detecting additive and dominant genetic effects of DNA methylation sites on quantitative traits and an application thereof, and belongs to the field of plant molecular breeding. The method comprises the following steps: 1) collecting population samples of germplasm resources, detecting phenotypic traits, and extracting genomic DNA from the samples; 2) constructing a bisulfite sequencing library by the genomic DNA of the samples, and sequencing; 3) identifying the DNA methylation sites from DNA methylation sequencing data, and genotyping; and 4) carrying outcorrelation analysis of the obtained genotyping data of the DNA methylation sites and the sample phenotypic trait data by a mixed linear model in Tassel 5.0 software package, and screening significantly correlated DNA methylation sites and analyzing the additive and dominant genetic effects. The method can provide new technical guidance for gene marker assisted breeding, and has important theoretical and breeding value.

The invention relates to the field of biomedicine, discloses a lung cancer recurrence prediction method based on DNA methylation of a tissue-specific enhancement subregion, and is used for solving the problem that an existing non-small cell lung cancer recurrence model is unreasonable in non-small cell lung cancer recurrence prediction. The method comprises the following steps: firstly, extracting DNA of a lung cancer patient, and carrying out DNA methylation detection to obtain a beta value of a DNA methylation site of a tissue specific enhancer region; wherein the DNA (deoxyribonucleic acid) methylation sites comprise chr1: 170667082, chr12: 85280518, chr17: 76561412, chr11: 9759297, chr15: 91915828, chr16: 1079990, chr2: 226797988, and chr7: 150477419; and then substituting the beta value of the DNA methylation site into a pre-constructed lung cancer recurrence model, calculating a model score, and obtaining a prediction result according to the model score and a plurality of score thresholds. The method is suitable for predicting the recurrence of the early non-small cell lung cancer.

The invention discloses a fluorescent biosensor for detecting DNA methylated transferase and preparation and application thereof. The fluorescent biosensor comprises a symmetric double-ring dumbbell and a 3D tetrahedral fluorescent scaffold, the symmetric double-ring dumbbell contains DNA methylation sites, methylation double strands can be formed under catalysis of DNA methylated transferase, enzyme digestion reaction is performed to form two single-chain dumbbell rings with consistent structures; the top ends of the 3D tetrahedral fluorescent scaffold contain the same hairpin and are modified with a fluorophore and a quenching group, the fluorophore and the quenching group are close to each other to cause fluorescence quenching, the sheared dumbbell ring can be combined with the hairpin to generate fluorescence, and DNA methylated transferase with different concentrations can generate fluorescence signals with different intensities through catalytic reaction. The DNA methylated transferase detection method established on the basis of the sensor is good in stability and specificity and high in sensitivity, can be used for screening MTase inhibitors, and has important significance on early detection and diagnosis of clinical cancers, drug research and the like.

The invention discloses a DNA methylation marker for Psoriatic Arthritis (PsA), a diagnostic reagent and application thereof. The DNA methylation marker comprises one or more of six methylation sites:chr14: 38061326, chr14: 38061320, chr9: 128585454, chr6: 37225002, chr3: 101901234 and chr6: 97369501. The specific DNA methylation sites are used as molecular markers for the first time to assist inscreening patients with Psoriatic Arthritis and Rheumatoid Arthritis (RA), and a new way and mode are opened up for diagnosis in the field. The DNA methylation sites are detected by using whole bloodof the patients, the purpose of early diagnosis of PsA is achieved, the required blood samples are few, the DNA methylation marker is convenient and easy to carry out, and the DNA methylation markerhas a good application prospect.

A prognosis risk assessment system for gastric cancer patients is characterized by comprising a gastric cancer methylation data acquisition module, a differential methylation site acquisition module and a prognosis model construction module. The gastric cancer methylation data acquisition module is used for acquiring TCGA gastric cancer methylation spectrum and a gastric cancer methylation spectrum data set GSE30601, the differential methylation site acquisition module is used for obtaining a plurality of hyper-methylation sites and a plurality of hypo-methylation sites, and the prognosis model construction module is used for constructing a 11-DNA methylation site risk scoring formula of each patient.

The invention belongs to the technical field of immunoassay, and discloses a methylation electrochemical immunoassay method, an electrode and an electrochemical sensor. The methylation electrochemical immunoassay method comprises the steps of: preparing a nanogold modified gold electrode, and preparing the electrochemical sensor; and immobilizing the anti-5-mC antibody and the enzyme-labeled secondary antibody, and performing electrochemical determination. The preparation method of the nanogold-modified gold electrode comprises the following steps: polishing a gold electrode with the diameter of 2mm into a mirror surface by using 0.05 [mu]m Al2O3 powder, and then carrying out ultrasonic cleaning for 5 minutes in ultrapure water, absolute ethyl alcohol and ultrapure water respectively; after drying at room temperature, putting the electrode into a prepared Piranha solution to activate for more than 15 minutes, then thoroughly cleaning with ultrapure water, and blow-drying with nitrogen; and immersing the treated gold electrode in a HAuCl4. 3H2O solution, and finally, fuly washing and cleaning the gold electrode with ultrapure water, and preparing the modified gold electrode. Along with the increase of the number of DNA methylation sites, the peak current of the DPV is increased.

The invention provides a DNA methylation marker for predicting the calf muscle dimension decline risk in weightless muscle atrophy. The DNA methylation marker comprises one or more of the following genes, and site annotation information thereof is shown in Table 3. The invention further provides a corresponding detection method. Excavation of the blood cell DNA methylation level and the calf muscle dimension finds that individual difference in human calf muscle dimension decline after weightlessness can be predicted through detection of the blood cell DNA methylation level, so that an individual which is more tolerant to a weightless condition is screened or calf muscle dimension change is scientifically researched. The invention provides a DNA methylation site set having a function of predicting the human calf muscle dimension decline risk after weightlessness.

The invention belongs to the technical field of biology, particularly discloses a time-of-flight mass spectrometry multi-target DNA methylation site quantitative detection method, and provides a high-throughput detection method of DNA methylation sites, which can design a specific probe for multiple methylation sites to be detected, and in the same reaction system, a quantitative methylation degree detection result is obtained. The method has the characteristics that the operation is simple, the methylation sites in different regions can be simultaneously detected, the corresponding experiment cost is reduced, and the like.

The invention provides a simple and convenient high-throughput DNA methylation detection method, which comprises the following steps: (1) carrying out oxidation treatment on a DNA sample by using strong base to oxidize cytosine in the DNA sample into uracil and oxidize methylated cytosine into thymine; (2) adding acid to neutralize strong base, and controlling the pH value of the reaction system within 6-8; and (3) carrying out library amplification and sequencing by adopting uracil-intolerant high-fidelity DNA polymerase. The invention develops a simple and convenient high-throughput DNA cytosine methylation detection technology which is named as DMA-seq. Compared with an existing DNA methylation detection kit, the DMA-seq has the advantages of being easy to operate, short in consumed time, small in DNA damage, good in data quality, high in methylation site targeting performance and the like, and is suitable for high-abundance DNA methylation site detection, especially in the early tumor screening direction.

The invention belongs to the field of biological genes technology, and discloses a DNA methylation site analysis method and a DNA biosensor. An AuNPs (at) rGO composite material is used for modifying a gold electrode; designing target sequences of a capture probe, an auxiliary probe and different methylation sites; performing DNA hybridization to induce signal amplification of gold nanoparticles aggregation; identifying a methylation site through anti-5methylcytosine antibody specifically; optimizing optimal experiment conditions of an electrochemical sensing analysis technology; performing electrochemical detection; and detecting the methylated DNA in the serum. According to the invention, a triple helix nucleic acid structure is combined with an electrochemical technology, a simple, stable and high-specificity DNA methylation site analysis technology is constructed, the maximum detection number of DNA methylation sites is 7, which is superior to results reported in other literatures, and a new technical means is provided for early clinical diagnosis of gene epigenetic disorder diseases and tumors and selection of a demethylation targeted therapy scheme.

The invention discloses a whole genome typing method based on DNA methylation site genotypes. The whole genome typing method comprises the following steps: step 1, collecting a sample; step 2, establishing a DNA library; step 3, sequencing the library; step 4, identifying a site; and step 5, genotyping. In the step 2, MBD treatment of the combined methylation sensitive restriction endonuclease refers to that the endonuclease of a non-to-be-detected area and the methylation sensitive restriction endonuclease are used for digesting DNA fragments at the same time, then fragments containing methylation areas are captured and reserved through an MBD column, and in the step 1, at different time periods such as daytime and night in a group, compared with an existing genotyping method, the method has the advantages that the influence of environments in different time periods on the DNA methylation state can be judged; according to the invention, the change of the DNA methylation state in different growth states can be judged; and according to the method, two different methods for treating DNA are adopted, so that the genotyping accuracy is improved.

The invention relates to the technical field of tumor molecular biology, and discloses a construction method of a dCas9-sgLINC00261 system. The method comprises the following steps: constructing a CRISPR-dCas9 pancreatic cancer cell line by using lentivirus, performing screening to obtain differential DNA methylation sites of a promoter region of LINC00261, constructing an sgRNA lentivirus vector,and transfecting the constructed sgRNA lentivirus vector into the CRISPR-dCas9 pancreatic cancer cell line. A dCas9-sgLINC00261 system is obtained. The invention also provides an application of the dCas9-sgLINC00261 system in preparation of drugs, markers or kits for preventing, detecting or treating pancreatic cancer. The cancer suppressor gene LINC00261 is modified through targeted demethylation, so that the endogenous expression quantity is increased, the pancreatic cancer proliferation and metastasis inhibiting capacity of the cancer suppressor gene is improved, and an innovative thoughtcan be provided for pancreatic cancer novel targeted drug research and development and clinical diagnosis and treatment.

The invention provides detection and application of methylation sites of human FITM1 and HIST1H2BJ genes. The invention discovers specific DNA methylation sites of genes related to primary hepatocellular carcinoma population, including the methylation site cg06186808 of the FITM1 gene and the methylation site cg04424621 of the HIST1H2BJ gene, and designs a pyrosequencing PCR primer and a pyrosequencing primer for the specific sites, and methylation of the specific sites can be quickly and accurately quantitatively detected through pyrosequencing.

The invention discloses human health and longevity markers based on specific peripheral blood DNA methylation sites and an application of the markers. The total number of the human health and longevity markers is 90. A single, or two or more markers can be used for predicting human health and longevity, and the human health and longevity markers have the characteristics of high accuracy and high sensitivity. The AUC value of 14 sites exceeds 0.85, and thus the human health and longevity markers can be well used for evaluating the potential of individual health and longevity.