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117 results about "Bisulfite salt" patented technology

Method for producing complex DNA methylation fingerprints

Method for characterizing, classifying and differentiating tissues and cell types, for predicting the behavior of tissues and groups of cells, and for identifying genes with changed expression. The method involves obtaining genomic DNA from a tissue sample, the genomic DNA subsequently being subjected to shearing, cleaved by means of a restriction endonuclease or not treated by either one of these methods. The base cytosine, but not 5-methylcytosine, from the thus-obtained genomic DNA is then converted into uracil by treatment with a bisulfite solution. Fractions of the thus-treated genomic DNA are then amplified using either very short or degenerated oligonucleotides or oligonuclcotides which are complementary to adaptor oligonucleotides that have been ligated to the ends of the cleaved DNA. The quantity of the remaining cytosines on the guanine-rich DNA strand and / or the quantity of guanines on the cytosine-rich DNA strand from the amplified fractions are then detected by hybridization or polymerase reaction, which quantities are such that the data generated thereby and automatically applied to a processing algorithm allow the drawing of conclusions concerning the phenotype of the sample material.
Owner:EPIGENOMICS AG

Reduced representation bisulfite sequencing with diversity adaptors

Described herein are methods, compositions and kits for the generation of bisulfite-converted libraries useful for conducting reduced representation bisulfite sequencing (RRBS). The methods described herein can be employed to generate RRBS libraries in a manner that is easier and more cost-efficient than conventional RRBS methods, and can be efficiently sequenced with next generation sequencing (NGS) techniques without the need for genomic, higher diversity sequencing controls such as PhiX spike-ins.
Owner:NUGEN TECH

Whole-genome methylation non-bisulfite sequencing library, construction and applications thereof

InactiveCN110820050ASolve the defect of low usage rateReduce usageNucleotide librariesMicrobiological testing/measurementDihydrouracilPhosphoric acid
The invention relates to the technical field of bioinformatics, particularly to a whole-genome methylation non-bisulfite sequencing library, a construction and applications thereof, wherein the sequencing library comprises a TET enzyme reaction solution, the TET enzyme reaction solution comprises the following independently packaged components: a TET enzyme oxidation buffer solution, and the TET enzyme oxidation buffer solution comprises, by micromole, (20-167)*10<3> parts of HEPES or Tris-Cl, (100-333)*10<3> parts of NaCl, 3.3*10<3> parts of alpha-KG or 2-oxoglutarate, 6.67*10<3> parts of ascorbic acid and 4*10<3> parts of adenosine triphosphate. According to the invention, by combining the kit, Fe(NH4)2(SO4)2 and TET enzyme, 5mc can be oxidized into 5cac, the 5cac is reduced into dihydrouracil under the action of a reducing agent, and T is identified through PCR sequencing, so that the the DNA methylation C-to-T conversion under the non-bisulfite condition is achieved, the defects ofbase imbalance and low sequencing data use rate of the existing methylation sequencing library constructed based on the bisulfite conversion are solved; and the formula further has effects of simplecomponents, extremely low TET enzyme use amount and significant cost reducing.
Owner:BEIJING GENEPLUS TECH +1

Kit for identification of gastric cancer and/or gastric polyps and application thereof

The invention relates to a kit for the identification of gastric cancer and / or gastric polyps and application thereof. The kit comprises a primer pair, wherein the primer pair is used for detecting the methylation level of a biological marker gene or corresponding segments in a biological sample of a tester; the primer pair is used for performing PCR (polymerase chain reaction) amplification reaction by using the biological marker gene or segments thereof treated by bisulfite as a template; the biological marker gene is selected from one or multiple of CDH1, DAPK, PAX5, RASSF1A, Reprimo, RNF180, RUNX3, SDC2, Septin9 and TCF4. The kit has the advantage that by jointly detecting the methylation level of the biological marker gene or corresponding segments, the sensitivity and specificity indetection of the gastric cancer and / or gastric polyps is improved, so that the correctness and reliability of the detection results are guaranteed, and a quick, reliable and accurate novel path is provided for the predicting, diagnosis and evaluation on the gastric cancer and / or gastric polyps.
Owner:BEIJING EXELLON MEDICAL TECH CO LTD

Integrated purification and measurement of DNA methylation and co-measurement of mutations and/or mRNA expression levels in an automated reaction cartridge

Methods of determining methylation of DNA are provided. In one illustrative, but non-limiting embodiment the method comprises i) contacting a biological sample comprising a nucleic acid to a first matrix material comprising a first column or filter where said matrix material binds and / or filters nucleic acids in said sample and thereby purifies the DNA; ii) eluting the bound DNA from the first matrix material and denaturing the DNA to produce eluted denatured DNA; iii) heating the eluted DNA in the presence of bi sulfite ions to produce a deaminated nucleic acid; iv) contacting said deaminated nucleic acid to a second matrix material comprising a second column to bind said deaminated nucleic acid to said second matrix material; v) desulphonating the bound deaminated nucleic acid and / or simultaneously eluting and desulphonating the nucleic acid by contacting the deaminated nucleic acid with an alkaline solution to produce a bi sulfite converted nucleic acid; vi) eluting said bi sulfite converted nucleic acid from said second matrix material; and vii) performing methylation specific PCR and / or nucleic acid sequencing, and / or high resolution melting analysis (HRM) on said bisulfite-converted nucleic acid to determine the methylation of said nucleic acid, wherein at least steps iv) through vi) are performed in a single reaction cartridge.
Owner:CEPHEID INC

High-throughput sequencing method for methylated DNA and use thereof

ActiveCN102971434BMicrobiological testing/measurementDNA preparationRepetitive SequencesMethylated DNA immunoprecipitation
The present invention provides a high-throughput sequencing method for methylated DNA, and use thereof. Particularly, the present invention provides a high-throughput sequencing method for methylated DNA, which combines methylated DNA immunoprecipitation, removal of repetitive sequences, and bisulfite treatment. The site of sequencing library will be decreased, and the cost will be reduced by using the method disclosed in the present invention.
Owner:INST OF PSYCHOLOGY CHINESE ACADEMY OF SCI +1

Method for simultaneously detecting genomic DNA polymorphism and methylation

The invention provides a method for simultaneously detecting genomic DNA polymorphism and methylation. The method comprises the following steps: performing DNA fragmentation, and digesting the 3' endof the DNA fragment with DNA exonuclease; adding a dNTPs mixture containing 5-hydroxymethylcytosine deoxyribonucleotide, and performing completion the 3' end of the DNA fragment; performing bisulfiteconversion treatment on the modified DNA fragment and connecting an adaptor adapted to a high-throughput sequencing platform, and finally constructing a sequencing library by PCR amplification. By combining a double-end sequencing mode of the high-throughput sequencing platform, genomic DNA polymorphism and methylation detection can be performed simultaneously, and the goal of simultaneous detection of genome and epigenome is achieved.
Owner:SHANGHAI EPIPROBE BIOTECH CO LTD
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