51 results about "Bisulfite" patented technology

The invention discloses a tetrahydrocarbazole enhanced fluorescent probe for detecting hydrosulfite as well as a preparation method and application of the tetrahydrocarbazole enhanced fluorescent probe. The fluorescent probe is a 2-(2-(5-(2, 2-dimethyl-2, 3, 4, 9-tetrahydro-1H-1, 3-methyl carbazole-6-yl) furan-2-yl) vinyl)-3-ethyl benzothiazole-3-onium iodised salt compound, namely, TC-FV-BT, and the fluorescent probe can be applied to the field of fluorescence detection. The probe TC-FV-BT can be used for specifically recognizing the hydrosulfite. Under sunlight, after bisulfite is added into a DMF / H2O (2: 8, v / v) solution of the probe TC-FV-BT, the color of the solution is changed from dark pink to white, under irradiation of ultraviolet light with the wavelength of 365 nm, the solution emits orange fluorescence at the emission wavelength of 605 nm, the fluorescence color of the solution is changed from colorless to orange, the response time is 4 min, the detection limit of the bisulfite reaches 77.27 nM, and the detection limit of the bisulfite reaches 77.27 nM. The fluorescent probe shows good selectivity and anti-interference performance, and has a good application prospect in the fields of environmental analysis, food and the like as a fluorescent probe for detecting hydrosulfite.

The invention belongs to the technical field of biological detection, and particularly relates to a bisulfite-free detection kit for specific site 5-methylcytosine and a detection method thereof. According to the method, a TAPbeta treatment method is adopted, 5hmC is sealed through beta-GT, 5mC oxidation is carried out through TET, and 5mC is reduced into dihydrouracil (DHU) through pyridine borane, so that the apparent difference between 5mC and C and 5hmC is converted into single nucleotide polymorphism difference. In addition, a flow microsphere technology based on PNA-assisted click chemical connection starting is established to detect DNA methylation specific sites. A PNA clip is innovatively introduced into click chemical connection, and non-specific connection is inhibited. Target sequence enrichment, signal amplification and signal acquisition are carried out through surface-functionalized magnetic nanoparticles, and the detection sensitivity is improved to fM. The invention provides a new tool for high-specificity and high-sensitivity epigenetic marker detection.

The present application relates to a method and system for inferring the age of an individual from semen or a semen stain, and a methylation detection kit, which comprises the following steps: firstly, extracting the genomic DNA of the semen or semen stain of the individual; secondly, performing bisulfite treatment on the genomic DNA; thirdly, selecting a plurality of sites in the CpG sites in the genomic DNA and amplifying the selected CpG sites to obtain the methylation rate of the selected CpG sites; and finally, performing regression analysis on the methylation rate of the selected CpG sites and the age of the individual to construct a regression model for inferring the age of an unknown individual from semen or a semen stain. The average absolute error of the present application reaches 1.68 to 4.44 years.

The application discloses a cervical cancer methylation detection kit and application thereof, the kit takes PAX5 gene, PAX1 gene, MICOS10-NBL1 gene and MICOS10 gene as detection targets; the kit mainly comprises specific primer probe dry powder combination, PCR reaction reagent dry powder, positive control product dry powder, negative control product dry powder and microdroplet generation oil. The sample is detected by bisulfite conversion, and the kit is combined with microdroplet digital PCR detection technology to realize accurate detection of the methylation level of the target gene. The detection verification result shows that the sensitivity of the kit for cervical cancer detection reaches 96.9%, the specificity reaches 95.7%, the kit has the advantages of accurate detection, simple operation and strong stability, can be widely applied to early screening, clinical auxiliary diagnosis and physical examination of healthy people of cervical cancer, and provides reliable technical support for early discovery and early treatment of cervical cancer.

The methods, compositions, and kits of the disclosure provide a novel approach for a whole genome, unbiased DNA analysis method that can be performed on limited amounts of DNA can be used to analyze DNA to determine its modification status. Aspects of the disclosure relate to a method for amplifying bisulfite-treated deoxyribonucleic acid (DNA) molecules comprising: (a) ligating an adaptor to the DNA molecules, wherein the adaptor comprises a RNA polymerase promoter comprising bisulfite-protected cytosines; (b) treating the ligated DNA molecules with bisulfite; (c) hybridizing the bisulfite-treated DNA molecules with a primer; (d) extending the hybridized primer to make double stranded DNA; and (e) in vitro transcribing the double-stranded DNA to make RNA.

The invention relates to a continuous preparation method of pentaerythritol bisulfite, which comprises the following steps: respectively and continuously introducing pentaerythritol emulsion and thionyl chloride into a continuous flow oscillation stirring reactor for continuous flow reaction, controlling the reaction temperature to be 40-90 DEG C, controlling the temperature to be 40-90 DEG C, and controlling the reaction time to be 1-3 hours; the stirring rotating speed is 200 to 800 r / min, the reaction pressure is 0.3 to 0.8 MPa, and the reaction time is 30 to 120 minutes. By using the continuous flow oscillation stirring reactor and specific reaction conditions, the continuous preparation is realized, and the pentaerythritol conversion rate, the product yield and purity, the reaction efficiency and the post-treatment simplicity are considered at the same time. The method has the remarkable advantages of simple process, low reaction operation difficulty, high reaction efficiency and the like, and can meet the comprehensive requirements of large-scale continuous industrial production on efficiency, cost and product quality.

The invention discloses an LAMP (loop-mediated isothermal amplification)-based rapid detection primer composition and kit for non-muscle invasive bladder cancer and application of the LAMP-based rapid detection primer composition and kit. The primer composition comprises a primer group for targeted amplification of a VIM gene and a TMEFF2 gene. According to the method, rapid nucleic acid extraction is achieved through a one-step method reagent, enzymatic methylation conversion is combined to replace a traditional bisulfite method, and nucleic acid damage and detection complexity can be remarkably reduced. A double-target detection system is constructed on the basis of VIM and TMEFF2 genes, and visual rapid detection of the non-muscle invasive bladder cancer is realized in combination with an LAMP detection technology. The whole detection process does not need a centrifugal machine or a PCR instrument, extraction, conversion and detection can be completed only through a metal bath or a water bath kettle, operation is easy and convenient, and cost is low. The kit provided by the invention is suitable for household self-test, reduces the detection burden of patients, and has wide clinical and household application prospects.

Methods and systems provided herein address current limitations of bisulfite-based methylation sequencing by improving the quality and accuracy of nucleic acid methylation sequencing and uses thereof for detection of disease. Methods that include minimally-destructive conversion methods for methylation sequencing as well as specialized UMI adapters provide for improved quality of sequencing libraries and sequencing information. Greater accuracy and more complete methylation-state information permits higher quality feature generation for use in machine learning models and classifier generation.

The invention relates to the technical field of molecular biological detection, in particular to a detection method for DNA methylation of a miR-34a gene promoter region and application. According to the method, the bisulfite reaction system is treated by two tubes, and the strategy of adjusting the reaction time according to the DNA dosage is combined, so that the conversion efficiency of the unmethylated cytosine is improved to 90% or above, and is obviously superior to that of the existing single-tube reaction.

This invention discloses a class of novel small-molecule fluorescent compounds based on thiazolyl-2-pyridone rings and their applications in analytical detection and diagnostics. The small-molecule fluorescent compounds of this invention have novel structures, readily available starting materials, mild synthesis conditions, and high yields. Some of the obtained compounds are responsive to amines, hydrazine, hypochlorite, and bisulfite, and are expected to serve as detection probes for these compounds. Other compounds exhibit long fluorescence emission and large Stokes shifts, effectively avoiding background spectral interference, generating Type I reactive oxygen species, and possessing near-infrared emission characteristics, showing potential as Type I photodynamic therapy photosensitizers.

A method for the continuous manufacture of taurine, comprising at least one of (a) – (c): (a) reacting an ethylene oxide and an alkali metal bisulfite to obtain a first intermediate product stream comprising an alkali metal isethionate; (b) reacting the alkali metal isethionate from the first intermediate product stream with ammonia to obtain a second intermediate product stream comprising an alkali metal taurate; and (c) converting the alkali metal taurate from the second intermediate product stream to obtain taurine and at least one by-product, wherein the by- product is suitable for reacting sulfur dioxide to alkali metal bisulfite, and separating the at least one by-product from the taurine to obtain at least one by-product stream comprising the at least one by-product and a product stream comprising the taurine; and an apparatus for the continuous manufacture of taurine.

The invention discloses a QPCR (Quantitative Polymerase Chain Reaction)-based DNA (Deoxyribose Nucleic Acid) methylation rate detection method, which comprises the following steps of: treating a DNA sample to be detected by bisulfite, dividing the DNA sample into a first sample and a second sample which are parallel to each other, and performing substrate amplification on a methylated DNA template and a non-methylated DNA template in the samples through PCR amplification by adopting the same amplification primer under the same condition; then respectively depleting dATP in a sample A reaction system and dGTP in a sample B reaction system through a template and a primer with thymine deoxyribonucleotide repetitive sequences or a template and a primer with cytosine deoxyribonucleotide repetitive sequences, and controlling independent detection of methylation and non-methylation templates through a d3TP system lacking dATP or dGTP; and finally, directly analyzing the fluorescence signal of the methylation template and the fluorescence signal of the non-methylation template, and calculating the methylation rate of the DNA sample to be detected through a methylation rate calculation function.

Methods of determining methylation of DNA are provided that include filtering a biological sample comprising a nucleic acid with a first matrix material to purify the DNA; eluting and denaturing the DNA to produce eluted denatured DNA; heating the DNA in the presence of bisulfite ions to produce deaminated nucleic acid; optionally contacting said deaminated nucleic acid with a second matrix material; desulphonating and eluting the bound deaminated nucleic acid; and performing methylation specific PCR, 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 multiple steps may be performed in a single reaction cartridge.

The present application belongs to the field of medicine and chemical industry, and relates to a method for purifying losartan. In the method, bisulfite or sulfite is used to remove color impurities of losartan, and the method has the advantages of mild treatment condition, short operation period, high decolorization efficiency and suitability for industrial production.

Provided are a method for preparing Bisulfite DNA and a use thereof. In the method, the process is simplified and optimized, and the same reagent is used as a reagent having both lysis and transformation functions, thereby avoiding the conversion of multiple systems, and reducing the steps of washing, elution and the like of a DNA sample.

The application provides a Septin9 methylation detection composition and application thereof, and the composition comprises a methylation-dependent restriction enzyme, a capture oligonucleotide, a universal primer and a Septin9 methylation-specific primer; the capture oligonucleotide comprises a first universal sequence, a folding sequence and a binding capture sequence from 5' end to 3' end in sequence; the folding sequence is at least partially identical to a 5' end sequence of a Septin9 methylation site after the methylation-dependent restriction enzyme is cut; and the binding capture sequence is specifically bound to a fragment region where the detected Septin9 methylation site is located. The application is based on a methylation-dependent restriction enzyme and a universal primer fluorescence quantitative PCR technology, does not need bisulfite conversion, and realizes accurate and specific Septin9 methylation detection.

The application discloses a methylation marker combination for detecting early esophageal cancer and application thereof. The marker combination comprises 178 methylation sites of 12 specific DNA regions and a specific internal marker region, and the methylation levels of the regions in plasma of patients with early esophageal cancer and healthy people are significantly different. The application also provides a primer combination and a kit for detecting the marker combination, and supports high-throughput deep sequencing. A detection system is used to extract plasma free DNA, perform bisulfite conversion and amplification, calculate a prediction value by using a logistic regression model, multiply the average methylation rate of each region by a corresponding weight coefficient, and compare the result with a threshold value to make a positive judgment. The application greatly improves the detection sensitivity of non-invasive screening for early esophageal cancer under the premise of ensuring high specificity, and has a wide clinical application prospect.

The methods, compositions, and kits of the disclosure provide a novel approach for a whole genome, unbiased DNA analysis method that can be performed on limited amounts of DNA can be used to analyze DNA to determine its modification status. Aspects of the disclosure relate to a method for amplifying bisulfite-treated deoxyribonucleic acid (DNA) molecules comprising: (a) ligating an adaptor to the DNA molecules, wherein the adaptor comprises a RNA polymerase promoter comprising bisulfite-protected cytosines; (b) treating the ligated DNA molecules with bisulfite; (c) hybridizing the bisulfite-treated DNA molecules with a primer; (d) extending the hybridized primer to make double stranded DNA; and (e) in vitro transcribing the double-stranded DNA to make RNA.

The present invention relates to a stabilized composition with significantly improved shelf-life stability. More specifically, the present invention relates to a stable liquid aqueous pharmaceutical composition comprising cyticine or a pharmaceutically acceptable salt thereof, at least one antioxidant selected from the group consisting of sulfite compounds, bisulfite compounds, and pyrosulfite compounds. The present invention also relates to the use of at least one antioxidant selected from the group consisting of sulfite compounds, bisulfite compounds, and pyrosulfite compounds for stabilizing a liquid aqueous pharmaceutical composition comprising cyticine.

The application discloses a solution containing natural macromolecule and a coagulation molding method. The fiber stream obtained by wet spinning of the solution containing natural macromolecule is coagulated and molded through at least one coagulation bath. The solution containing natural macromolecule contains metal hydroxide and an additive selected from at least one of urea, zinc oxide and thiourea. Bicarbonate or bisulfite or a combination of bicarbonate and carbonate or a combination of bisulfite and sulfite is used as a component of the coagulation bath. The low-cost, green, efficient, non-toxic, harmless and inexpensive inorganic salt solution does not contain strong acid and organic solvent, and can be used to prepare long fibers with circular cross section and excellent mechanical properties. The long fibers have smooth surface, softness and luster. In the coagulation bath, the bicarbonate and carbonate or the bisulfite and sulfite can be easily separated and recovered.

The invention belongs to the field of new materials, and discloses a preparation method of 1-propylene-1, 3-sultone, which comprises the following steps: step 1, addition reaction: using propargyl alcohol and alkali metal sulfite and / or alkali metal hydrosulfite as raw materials, and carrying out addition reaction in the presence of a catalyst and a free radical initiator to obtain an addition product; step 2, acidification reaction: adjusting the pH value of the addition product to 4 or below 4, and separating to obtain an acidified product; step 3, cyclization reaction: carrying out cyclization reaction on the acidified product to obtain 1-propylene-1, 3-sultone; the catalyst is soluble metal salt which is soluble in water. According to the method, the catalyst and the free radical initiator which are low in price are compounded, so that a product with relatively good conversion rate and selectivity is obtained.

The methods, compositions, and kits of the present disclosure provide a new method for whole genome, i.e., unbiased DNA analysis, that can be performed on limited amounts of DNA. It can be used to analyze DNA to determine its modification status. Aspects of the present disclosure relate to methods for amplifying a bisulfite treated deoxyribonucleic acid (DNA) molecule, the method comprising: (a) ligating an adaptor to the DNA molecule, wherein the adaptor comprises a RNA polymerase promoter comprising bisulfite protected cytosines; (b) treating the ligated DNA molecule with bisulfite; (c) hybridizing the bisulfite treated DNA molecule to a primer; (d) extending the hybridized primer to produce double stranded DNA; and (e) in vitro transcribing the double stranded DNA to produce RNA.

The invention discloses a cellulose acetate based 1, 8-naphthalimide fluorescent probe for detecting hydrogen sulfite (HSO3 <->) as well as a preparation method and application of the cellulose acetate based 1, 8-naphthalimide fluorescent probe. The preparation method comprises the following steps: by taking 4-bromo-1, 8-naphthalic anhydride as a raw material, carrying out condensation reaction on the 4-bromo-1, 8-naphthalic anhydride and 4-aminobutyric acid to prepare 4-(4-bromo-1, 8-naphthalic diimide) butyric acid, namely BNA-B; the preparation method comprises the following steps: carrying out coupling reaction on BNA-B and 4-formyl phenylboronic acid to prepare 4-(4-(4-formyl phenyl)-1, 8-naphthalimidoyl) butyric acid, namely FNA-B; the preparation method comprises the following steps: carrying out aldol condensation reaction on FNA-B and 1, 2, 3, 3-tetramethyl-3H-indole iodide to prepare a compound INA-B; and the compound INA-B and cellulose acetate (CA) are subjected to an esterification reaction, such that the cellulose acetate-based 1, 8-naphthalimide fluorescent probe INA-B-CA is prepared. The compound INA-B-CA can specifically recognize HSO3 <->, under the irradiation of 365nm ultraviolet light, after HSO3 <-> is added into a DMSO / PBS solution of INA-B-CA, the fluorescence color of the solution is changed from yellow to blue, and the probe has a good application prospect.

This invention discloses a method for detecting 5-hydroxymethylcytosine based on triple cascade signal amplification, belonging to the field of biochemical detection technology. The method includes the following steps: 1) preparation of 5hmC-specific glycosylation and azide labeling; 2) magnetic bead capture and enrichment separation of target DNA; 3) TdT-mediated nucleic acid end polymerization and extension; 4) APE1-triggered primer release and rolling circle amplification; 5) DNase-catalyzed cleavage and fluorescence signal detection. This invention is bisulfite-free and sequence-independent, achieving extremely low background and ultra-high sensitivity detection with a detection limit of 4.41 fM. It can be widely applied to early clinical cancer screening, biomarker detection for neurological or cardiovascular diseases, and provides a powerful platform for low-abundance epigenetic analysis in clinical diagnosis and biomedical research.

The application discloses a fluorescent probe for dual-channel distinguishing detection of hypochlorous acid and bisulfite in ferroptosis, a preparation method and application thereof, and belongs to the technical field of fluorescent probe detection and analysis of HClO and HSO3 ‑ . The fluorescent probe is (E)-3-ethyl-1,1-dimethyl-2-(4-(methylthio)styryl)-1H-benzo[e]indol-3-ium iodide, and a structural formula thereof is shown in the description; the application further discloses a preparation method of the fluorescent probe and application of the fluorescent probe in simultaneous, rapid and high-selectivity identification of HClO and HSO3 ‑ and preparation of imaging detection reagents of HClO and HSO3 ‑ in organisms. The fluorescent probe can realize dual-channel distinguishing detection of HClO and HSO3 ‑ in pure PBS solution, has fast response, low detection limit and high selectivity, and has good imaging effect on endogenous HClO and HSO3 ‑ generated in the ferroptosis process.

The application relates to the field of biomedical detection technology, in particular to a bladder cancer postoperative recurrence monitoring method based on a ctDNA methylation spectrum, which comprises the following steps: collecting postoperative patient cell-free plasma and extracting circulating tumor DNA; performing bisulfite conversion treatment on the DNA; performing targeted amplification using a multiplex PCR primer group designed for a group of predetermined genomic methylation regions related to recurrence, constructing a sequencing library; performing high-throughput sequencing on the library, obtaining methylation level data of a plurality of CpG sites to form a sample methylation spectrum data matrix; the core of the application is that tumor heterogeneity is overcome through multi-marker combination detection, and a machine learning model is used to integrate multidimensional data to realize precise risk stratification, which has the advantages of non-invasiveness, high sensitivity, high specificity and quantifiable output, and provides an effective tool for individualized follow-up management of bladder cancer postoperation.

The application discloses a preparation and application of a near-infrared ratio type bisulfite fluorescent probe, and relates to the technical field of bisulfite fluorescent probes. The probe comprises a near-infrared compound, and a structural formula is as follows: the preparation method and the application of the compound are also provided. The bisulfite ratio fluorescent probe can be reacted with bisulfite to generate a change in a fluorescence spectrum, so that quantitative detection of the bisulfite can be realized. The probe is very sensitive to the reaction with the bisulfite, so that the detection of the bisulfite is facilitated. The probe has good stability, and can be used for a long time. The bisulfite fluorescent probe is a high-selectivity and rapid-response bisulfite fluorescent probe, and is simple to synthesize.

This invention provides an SDC2 methylation detection kit and its application. The kit includes a methylation-dependent restriction endonuclease, a capture oligonucleotide, and universal primers. The capture oligonucleotide includes a first universal sequence, a folded sequence, and a binding capture sequence from the 5' end to the 3' end. The folded sequence is at least partially identical to the 5' end sequence of the SDC2 methylation site after digestion with the methylation-dependent restriction endonuclease. The binding capture sequence specifically binds to the fragment region containing the detected SDC2 methylation site. This invention, based on methylation-dependent restriction endonuclease and universal primers, utilizes real-time quantitative PCR technology, eliminating the need for bisulfite conversion, and achieves sensitive and specific detection of SDC2 methylation with a detection limit as low as 10 copies / reaction.