177 results about "Molecular hybridization" patented technology

The present invention provides binding assays, referred to here as fluorescence proximity assays or FPA. The inventions detect binding of target molecules in a sample to a molecular probe or probes that specifically bind or hybridize to those molecules. In particular, the molecular probes are immobilized to a bead or particle, such as colloidal gold, the reflects fluorescent energy from a fluorophore. The derivatized beads are contacted to a sample of fluorescently labeled target molecules, and binding of the target is indicated by an increase in the fluorescent signal. Kits are also provided that contain materials and reagents to performing a fluorescence proximity assay.

The invention provides a preparation method for an in situ self-assembled organic / inorganic hybrid membrane based on coordination, and belongs to the technical field of membrane separation. The method provided by the invention comprises the following steps: preprocessing the organic porous membrane and enabling the surface of which to be charged; preparing membrane casting solution by dissolving metal ion, organic ligand and polymer in a solution, and carrying out standing of the membrane casting solution for deaeration; conducting dynamic filtering or static deposition of the membrane casting solution on the surface of the organic porous membrane in an alternating manner for a period of time, so as to enable the metal ion and the organic ligand to generate hybrid particle on the surface of the membrane through the layer upon layer self-assembly method in the presence of polyelectrolyte, and forming an ultra-thin separation layer which is uniform in dispersity, high in loading capacity, and can realize molecular hybridization. The invention provides a novel preparation method for the organic / inorganic hybrid membrane, and nanofiltration membrane prepared by the method is provided with the advantages of high reject rate, big flux and the like, and the method can be widely used in the filed of water processing.

The method is performed by means of a pair of electrodes that are arranged at a distance and within a sensing zone. A nucleic acid capture molecule with an uncharged backbone and a nucleotide sequence that is at least partially complementary to at least a portion of a strand of the target nucleic acid molecule, is immobilised on an immobilisation unit. The immobilisation unit, which is arranged within the sensing zone, is contacted with a solution suspected to comprise the target nucleic acid molecule, which hybridizes to the nucleic acid capture molecule. An activation agent is added, which has an electrostatic net charge complementary to the net charge of the target nucleic acid molecule. It associates to the complex of nucleic acid capture molecule and target nucleic acid molecule. Added is a water soluble polymer with at least one polymer strand and with an electrostatic net charge that is complementary to the net charge of the activation agent. Thus the polymer associates to the activation agent. A metal salt is added, which can act as an oxidant and the metal ions of which have an electrostatic net charge complementary to the net charge of the polymer. The metal ions associate to the polymer. Upon adding a reducing agent, the latter reduces the metal ions, forming a metal wire. The presence of the analyte molecule is determined based on an electrical characteristic of a region in the sensing zone that is affected by the metal wire.

The invention provides a kit for separating genome DNA by using magnetic balls and application thereof. The kit comprises magnetic balls, a magnetic frame, a genome DNA extraction reagent (lysing solution, binding solution, rinsing solution A, rinsing solution B and eluent) and specifications; the main steps of extracting the gene DNA comprise cell lysis, nucleic acid absorption, impurity removing and nucleic acid elution. The application of the kit for extracting the genome DNA does not need to use large-toxicity organic solvents of phenol, chloroform and the like, has good safety, and simple, fast and time-saving operation, and simultaneously can extract a plurality of samples. The kit can extract the genome DNA from materials of animals, plants, bacteria, fungus, blood, virus, animal source feed stuff, samples in the forensic medicine and the like, the DNA has high yield and purity, and the obtained genome DNA can be used for experiments such as PCR amplification, gene cloning, construction of genomic library, sequence measurement, molecular hybridization, molecular marking and the like. The kit can be stored at the temperature of 4 DEG C, also can be placed at room temperature, and is convenient to transport.

Disclosed herein include systems, methods, compositions, and kits for molecular barcoding on the 5′-end of a nucleic acid target. After barcoding a nucleic acid target using an oligonucleotide barcode comprising a target binding region and a molecular label to generate a barcoded nucleic acid molecule, an oligonucleotide comprising a complement of the target binding region can be added to generate a barcoded nucleic acid molecule comprising the target-binding region and the complement of the target-binding region. A stem loop is formed with intra-molecular hybridization of the barcoded nucleic acid molecule, which can be extended to generate an extended barcoded nucleic acid molecule comprising the molecular label and a complement of the molecular label.

Devices, systems, and methods for detecting nucleic acid hybridization, including single nucleic base mutations at low concentrations, are disclosed, using capture units having nanoparticles with attached single-stranded oligonucleotides that are capable of hybridizing target oligonucleotides and reporter molecules having nanoparticles with attached single-stranded oligonucleotides, without the use of labeling or target modification.

The invention relates to a method for detection Listeria monocytogenes with loop-mediated isothermal amplification-CdS nanoparticles labeling electrochemistry. The method is characterized in that the method comprises the following steps of: obtaining double-stranded target sequence of amplification reagent-Listeria monocytogenes to be detected by means of loop-mediated isothermal amplification reaction; pyrolyzing a double-stranded target sequence in water bath into a single-stranded target sequence, then self assembling and fixing at the surface of gold electrode, so that the target sequence modified electrode of the Listeria monocytogenes to be detected is obtained; afterwards, labeling the probe sequence coming from the Listeria monocytogenes by CdS nanoparticles to obtain a labeled probe sequence; carrying out molecular hybridization on the labeled probe sequence and the target sequence of the target sequence modified electrode, and carrying out electrochemical detection by simultaneity plating Hg anodic stripping method; detecting out cadmium ions corresponding to the species-specific genes (actA) of the detected Listeria monocytogenes. The method has the advantages of speediness, strong specificity, high sensitivity and convenient use.

The invention discloses a method which is used for detecting and quantifying target material and based on nucleic acid aptamer and a solid-phase biosensor used for detecting, wherein, the method comprises the following steps: (1) capture probes are connected to the surface of magnetic particle, and the sequence of the capture probes comprises the nucleic acid aptamer sequence of the target material; (2) a sample to be tested is added for reaction; (3) signal probes and bridge molecules are hybridized to obtain double-stranded nucleic acids, and the signal probes are added into the reaction system obtained by step (2) through a double-stranded mode for reaction, the magnetic separation is then carried out to remove free signal probes; (4) the signal probes are separated from the surface of the magnetic particle by a nucleic acid denaturation way, and the fluorescent signals on the signal probes are detected. The method is suitable for detecting wide target materials comprising protein, organic micromolecule, metallic ions, and the like. The method not only has higher sensitivity, but also solves the nonspecific problem and has important meanings in bioinstrumentation, early diagnosis and treatment of disease.

The present invention is directed to devices and methods for molecular association, particularly to devices and methods for hybridization of nucleic acids utilizing temperature gradients and imaging thereof. In one aspect, a molecular hybridization system generally includes a substrate having a plurality of molecular probes attached thereto, the plurality of probes being generally present in multiple copies arranged in localized formations on the surface of the substrate. The molecular hybridization system further generally includes a chamber that encloses the plurality of molecular probes such that a fluid containing sample may be applied and kept in contact with the substrate having the probes thereon. The molecular hybridization system also includes a temperature affecting system that generally produces at least one desired temperature on the surface of the substrate and in the adjacent fluid within the chamber.

A unimolecular probe for hybridization to a molecule comprising a target nucleic acid sequence, the probe includes: a first nucleic acid sequence complementary to the target sequence (target-binding sequence); a second nucleic acid sequence complementary to a portion of the first nucleic acid sequence and capable of hybridization therewith to form a first intramolecular duplex. In use, the target and target-binding sequence hybridize to form a duplex. A probe can be used to detect a molecule containing the target sequence, act as a primer for synthesis or amplification, etc.

The present invention is directed to devices and methods for molecular association, particularly to devices and methods for hybridization of nucleic acids utilizing temperature gradients and imaging thereof. In one aspect, a molecular hybridization system generally includes a substrate having a plurality of molecular probes attached thereto, the plurality of probes being generally present in multiple copies arranged in localized formations on the surface of the substrate. The molecular hybridization system further generally includes a chamber that encloses the plurality of molecular probes such that a fluid containing sample may be applied and kept in contact with the substrate having the probes thereon. The molecular hybridization system also includes a temperature affecting system that generally produces at least one desired temperature on the surface of the substrate and in the adjacent fluid within the chamber.

The invention provides an electrochemical DNA biosensor for detecting the BCR / ABL fusion gene of chronic myeloid leukemia (CML), which comprises the following steps of: (1) designing and synthesizing the specificity sequence of the CML according to the fusion locus of the universal primer sequence of the selected CML gene type to be detected; and (2) building the electrochemical DNA biosensor by combining a chemical bonding technology, a molecular hybridization technology, a locked nucleic acid probe technology with an electrochemical enzyme linked immunity technology for detecting the BCR / ABL fusion gene of the CML. The biosensor greatly strengthens the sensitivity and the specificity, thereby being capable of realizing the early diagnosis of the chronic myeloid leukemia (CML).

The invention discloses a method and a kit for rapidly constructing a target nucleic acidsequencing library on the basis of probe capture and enrichment. The method comprises following steps and adopts reagents required for the steps: genome DNA is subjected to fragmentation; DNA end repair is performed; a probe and a single-stranded DNA fragment formed through thermal denaturation are subjected to molecular hybridization; biotin-avidinmagnetic bead separation is executed; the probe is taken as a primer for primer extension, and a complementary strand is synthesized; one end of a double-stranded DNA fragment is connected with an linker, and the other end is not connected with the linker; PCR amplification is performed; DNA fragment lengthselection is executed; the reagents are used for completing the reactions of all the steps. Compared withconventionaltarget nucleic acid enrichment technologies, the method has the advantages of good fidelity, good reproducibility, high speed and high universality.

The invention discloses a formula and a preparation method of a POSS-based regenerative medical material. The formula comprises 38-80 parts of trialkoxysilane, 40-72 parts of methanol, 10-22 parts deionized water, 2.6-3.2 parts of concentrated hydrochloric acid, 5-20 parts of phytic acid and 23-36 parts of calcium-containing salt, and a calcium and phosphorus element inlaid novel bioactive material with a cage structure composed of a Si-O- Si bond framework of silicon oxide is formed by adopting a specific proportion and accordingly achieves higher bioactivity. The material can achieve molecular hybridization enhancement with other medical macromolecules; The active groups on the periphery of the cage structure can be grafted with target functional groups so as to realize targeted repair.

The invention relates to a method for detecting a product of loop-mediated isothermal amplification (LAMP) by gold nano-particles. The method comprises the following steps: firstly, preparing gold nano-particles labeled with molecular probes; amplifying DNA or RNA to be detected by the loop-mediated isothermal amplification; and molecularly hybridizing the DNA molecules of the product of looped-mediated isothermal amplification by the gold nano-particles labeled by the molecular probes so as to promote the gold nano-particles to be close and cause color changes. According to changes of color signals, the method determines whether a sample to be detected contains the DNA or the RNA to be detected or not. The method has the advantages of high sensitivity, good specificity, simple and convenient operation, quickness without professional equipment and the like.

The invention discloses a surface plasma resonance probe with a silver-gold core-satellite structure and a preparation method thereof. The preparation method comprises the steps of: firstly, sequentially and quickly adding a glucose solution and a hexadecyl trimethyl ammonium bromide solution into a quantitative silver ammonia solution for reacting to prepare a nano silver cube solution; and then, standing a clean glass substrate in the nano silver cube solution to prepare a nano silver cube glass substrate; fixing single stranded DNA molecules on the prepared nano silver cube glass substrate; hybridizing the single stranded DNA with a miRNA molecule; and finally, preparing gold nanoparticles and decorating the gold nanoparticles at one end of the miRNA molecule to prepare the surface plasma resonance probe with a silver-gold core-satellite structure and capable of being applied in early detection of lung cancer. The particle diameter of the prepared surface plasma resonance probe is 50-70nm, the scattered spectrum peak is located at 450-650nm and the detection sensitivity is as high as 10nM.

The invention relates to a gene chip and a method; aiming at specific identification target genes of fourteen common pathogenic bacteria, the invention designs a corresponding primer and probe, and constructs a composite gene chip; the primer is marked by biotin, then is used for amplification of a target gene fragment of the substance to be detected by a PCR method; the amplification product is hybridized with the probe on the composite gene chip; through dual specific detection of the primer and the probe, the compound deposits on the composite gene surface, and specific molecules is hybridized to be converted into a visual signal; therefore, various toxigenic microorganisms can be detected. The invention can detect 14 common pathogenic bacteria, is high in detection flux, strong in specificity, good in sensitivity, rapid and effective.

The invention relates to a primer, a probe and a method for detecting entomophily or contact transmission pathogens by using a liquid phase chip, which are used for detecting nine clinical common entomophily or contact transmission infectious disease pathogens. The invention can detect the nine clinical common entomophily or contact transmission infectious disease pathogens based on an MASA (multi-analyte suspension array) liquid phase chip technology, homology analysis is performed respectively according to all nucleotide sequences of 9 target viruses which can be retrieved in a gene bank mainly, the degenerate primer and the specific probe are designed, two turns of PCR (polymerase chain reaction) and molecular hybridization are further performed, and a Luminex100 system is further usedfor detection, thereby determining types of the pathogens contained in the sample. The invention is most important for adopting a correct treatment scheme and timely taking measures for preventing disease transmission by detection and early diagnosis of the 9 clinical common entomophily or contact transmission infectious disease pathogens. The invention has the advantages of fast detection speed,simpleness in operation, high sensitivity, good specificity and the like, and is conductive to popularization.

The invention discloses a nested asymmetric PCR reagent kit for detecting alpha 2 globin gene point mutation. The reagent kit comprises a nested asymmetric PCR amplification primer, a fluorescent probe, a quenching probe, an LADNA polymerase and the like. The alpha-thalassemia point mutation genotype of a detected sample is detected by virtue of specific amplification of five point mutation destination fragments WS, QS, CS, CD30 and CD31 of the human alpha 2 globin gene and molecular hybridization and unwinding analysis of a specific fluorescent probe. The reagent kit has good sensitivity and accuracy to wild type and mutation type of alpha-thalassemia five point mutation positions WS, QS, CS, CD30 and CD31, has good repeatability and stability, and is totally suitable for clinical detection of alpha-thalassemia point mutation.

The invention relates to a xylanase mutant with improved specific enzymatic activity and a coding gene and application thereof. The xylanase mutant a protein shown in (a), (b) or (c): (a), a protein composed of amino acid sequences shown in SEQ ID NO. 1; a protein derived from (a) by subjecting the amino acid sequences shown in SEQ ID NO. 1 to substitution, deletion and / or addition of one or more amino acid residues and which has xylanase activity; (c), a protein coded by subjecting the amino acid sequences of code (a) and the amino acid sequences of code (b) to molecular hybridization and which has xylanase active amino acid sequences. The xylanase mutant according to the invention has specific enzymatic activity increased by 2.8 times, is applicable to degrading xylan substrates, has a wide range of acting temperature and pH, and has the advantages such as good resistance to acids and bases.

The invention discloses a method for preparing a conjugated molecular hybridization semiconductor photocatalytic material by using mechanochemistry, and belongs to the field of inorganic materials. The method comprises the following steps of uniformly mixing molecules containing conjugated large Pi bonds and an inorganic semiconductor nanomaterial according to a certain mass proportion; adding the mixture in a high-energy ball milling tank under the condition of a certain ball / material ratio; and treating the mixture for a certain time at proper ball milling revolving speed to obtain the conjugated molecular hybridized semiconductor catalyst material. The method has the advantages that a technological process is simple, the method is easy and convenient to operate, solvent is not used, aftertreatment is simple, preparation cost is low, the prepared composite catalyst material is high in activity, and the method is suitable for preparing the conjugated molecular hybridization modified semiconductor photocatalytic material in batches.

The invention discloses a streptomycete constitutive promoter and applications thereof, and provides an DNA fragment. The DNA fragment is any one selected from the following DNA molecules in 1)-3): 1) an DNA molecule represented by the sequence 1 in a sequence table; 2) an DNA molecule with promoter functions and hybrid with the DNA molecule shown in 1) under strict conditions; 3) an DNA molecule with more than 90% of homology with the DNA molecule shown in 1) and with promoter functions. The experiments shows that a strong constitutive promoter kasO*p is screened out, and the promoter and the erythromycin promoter ermE*p are compared in representation of transcriptional level and expression level in streptomyces coelicolor, streptomyces avermitilis and streptomyces venezuelae, and the results show that the modified promoter kasO*p is better than the the erythromycin promoter ermE*p.

The invention provides a high-throughput low-cost SNP (single nucleotide polymorphism) genotyping method based on a liquid molecular hybridization principle. The method comprises the following steps: extracting biological genome DNA (deoxyribonucleic acid) and carrying out biotin labeling on the biological genome DNA; designing site-specific hybridization primers LSP1 and LSP2 and carrying out 5' phosphorylation on LSP2; hybridizing an LSP1 mixture and an LSP2 mixture with the genome DNA to obtain a hybridization adsorption product; carrying out extended linkage reaction to obtain a target DNA fragment; carrying out a round of PCR (polymerase chain reaction) amplification on a universal primer; carrying out PCR amplification on the Barcode specific primer of the recovered target fragment; carrying out high-throughput sequencing; obtaining SNP site genotyping information through analysis. The method combines the site selection flexibility of the liquid hybridization technology with the advantages of high throughput and low cost of the high-throughput sequencing technology and has great application value and wide popularization prospect in the research fields such as large-scale screening of SNP, genome-wide association study, population diversity evaluation, gene function analysis and the like.

The invention belongs to the technical field of molecular biology, in particular to a nucleic acid releasing agent and a method and application for quickly releasing nucleic acid. The nucleic acid releasing agent is prepared from 5-500 mM of Tris-HCl, 50-1000 mM of NaCl, 0.1-10 mM of EDTA, SDS with the mass to volume ratio being 0.01-2%, LLS with the mass to volume ratio being 0.04-3% and betaine with the mass to volume ratio being 0.01-2%. According to the method for quickly releasing nucleic acid, nucleic acid in serum, plasma, urine, saliva, oral swabs, dried blood stains, forensic medicine samples and environmental samples can be quickly released, and the released nucleic acid can be applied to downstream experiments such as detection, clone, sequence analysis and molecular hybridization.

The invention relates to the field of estimating a microbial count in a sample using molecular techniques. The invention furthermore relates to a kit for performing such estimation and use of the estimation method for providing bacterial cell counts in milk and for diagnosing infectious conditions in animals and human beings. Furthermore, the invention relates to a method for quality control of food (especially milk and dairy products), feed and water samples using molecular techniques. Reproducible and reliable estimates of a microbial count can be obtained using a molecular hybridisation assay between a target nucleic acid sequence and a labelled probe. Proper calibration and quantitative recording of the signal produced by the label can estimate a microbial count. One important feature of the invention is the use of locked nucleic acid (LNA) monomers in the probes.

The invention discloses a telomere length detecting method based on a biological nano channel of aerolysin. The method comprises the following steps: (1) opening the G-quadruplet structure of telomere DNA of human; (2) preparing a biological nano channel of aerolysin: (a) activating aerolysin; (b) preparing a phospholipid n-decane solution; (c) preparing phospholipid bilayer; (d) forming a biological nano channel of aerolysin; (3) using the biological nano channel of aerolysin to detect the telomere length: (a) collecting monomolecular signal data; (2) analyzing the monomolecular signal data. Compared with the prior art, the provided method has the advantages that the operation is simple and efficient; enzyme digestion, molecular hybridization, or fluorescence labeling is not needed; the consumption of DNA substrate is little; the detection sensitivity and resolution are high; the price is cheap; the absolute length of telomere can be precisely and directly measured in real time; and the method has a positive meaning for bioscience research.

The invention discloses a bacteria detecting technology which comprises designing special probes with biological information, including designing 34 pieces oligonucleotide as filtered probes and the corresponding PCR and hybridizing reacting conditions; enlarging via PCR and marking bacteria 23srRNA gene internal sub array with Digoxin, and hybridizing the PCR products and a group of special oligonucleotide probes; after enzyme-linked immunoassay color-rendering, identifying whether has aquatic infectious pathogeny bacteria from the sample.

The invention discloses a microcystin test chip. The microcystin test chip comprises a substrate and probes combined on the substrate, wherein the probes are six specific probes related to six conserved region sequences in a microcystin synthetic gene cluster. The invention also discloses a kit provided with the test chip and a method for detecting microcystin by utilizing the kit. Compared with the prior art, the invention has the advantages that: a biochip and a molecular hybridization technology are combined, the fussy polymerase chain reaction (PCR) amplification is rejected, the detection time is shortened, and the high-throughput and high-sensitivity rapid detection can be performed.

Devices, systems, and methods for detecting nucleic acid hybridization, including single nucleic base mutations at low concentrations, are disclosed, using capture units having nanoparticles with attached single-stranded oligonucleotides that are capable of hybridizing target oligonucleotides and reporter molecules having nanoparticles with attached single-stranded oligonucleotides, without the use of labeling or target modification.