132results about How to "Improve hybridization efficiency" patented technology

The present invention relates to a process for detecting the presence of at least one specific nucleic acid sequence in a sample containing a nucleic acid or a mixture of nucleic acids by amplifying the nucleic acid using polymerase chain reaction, cleaving the amplified products with uracil DNA glycosylase to obtain short DNA segments and detecting the DNA fragments by using reverse blot hybridization.

The invention relates to a microfluidic device with microchannels that have separated regions which have a member of a specific binding pair member such as DNA or RNA bound to porous polymer, beads or structures fabricated into the microchannel. The microchannels of the invention are fabricated from plastic and are operatively associated with a fluid propelling component and detector.

The invention relates to methods and systems for use of bio-discs in quantifying an amount or concentration of one or more substances that are present in a biological sample. The system includes an optical disc having one or more samples deposited thereon, an optical element configured to emit and divert radiation onto the samples and components for measuring spectral change of the compound and determining an amount or concentration of one or more target substances. The bio-disc is prepared using a method including providing a membrane that is or can be dimensional to fit within a channel of the bio-disc, applying a reagent on the membrane which is configured to allow the reagent to be released from the membrane into a solution in contact with the membrane, and depositing the membrane in a channel of the bio-disc.

A method of using Neutrilized DNA (N-DNA) as a surface probe for a high throughput detection platform is disclosed. FET and SPRi are used as high throughput detection platforms to demonstrate that the N-DNA surface probe produces good results and enhances detection sensitivity. The N-DNA modifies the charged oxygen ions (O−) on the phosphate backbone through methylation, ethylation, propylation, or alkylation, so that the backbone is not charged after this modification to increase the hybridization efficiency, sensitivity and to make the signal more clear.

The present invention relates to methods and systems for the detection of specific sequences of nucleic acids or oligonucleotides. It relates more particularly to a conjugated enzyme based assay system utilizing reflective and / or transmissive optical bio-discs for detection of specific sequences of nucleic acids. In the assay according to the present invention, an enzyme reaction is used to detect the presence of an analyte (DNA or RNA) in a microchannel in an optical bio-disc. The analyte is immobilized by hybridization with a specific capture probe on a capture layer on the surface and the signal that is generated is localized and specific. The signal can be in the form of a pellet, a fluorescent product, and / or a colored product, and can be detected and quantified by an optical bio-disc reader utilized in conjunction with the inventions hereof. This assay is thus quantitative in nature.

The invention relates to a method for using a DNA tetrahedron as a scaffold on a nano-particle surface and initiating rolling circle amplification reaction; after the prepared DNA tetrahedron is assembled on the nano-particle surface, a section of single-stranded DNA extending on the DNA tetrahedron can be used as primer DNA; circular DNA complementary with a special aptamer sequence is used as an amplification template; the circular DNA is hybridized with the primer DNA while the rolling circle amplification reaction is initiated; and the prepared product is multiple long single-stranded DNA composed of multiple apatmer structures on the nano-particle surface. The method disclosed by the invention is capable of effectively regulating and controlling the DNA distribution density on the nano-particle surface, so that the hybridization efficiency of the primer DNA and the circular DNA is increased; by means of long single-stranded DNA composed of the multiple aptamer structures prepared through the rolling circle amplification technology, the CTC (Carbon Tetra Choride) capture efficiency is greatly increased; and thus, the method disclosed by the invention has potential application value in the research fields, such as cell detection, cell capture, cell imaging and the like.

The invention provides oligonucleotides, kits and methods for genotyping HPV.

The present invention discloses a DNA biosensor, which comprises an organic thin film transistor and a DNA molecule probe, wherein the DNA molecule probe is adopted as a recognition component, the organic thin film transistor is adopted as a sensor transducer for converting a biological signal detected by the DNA molecule probe into an easily detected electric signal, the organic thin film transistor comprises a base plate substrate, a gate electrode, an insulating layer, organic semiconductor layers and source drain electrodes, and the organic semiconductor layer forms an active layer or a carrier injection layer, such that the DNA molecule probe is fixed on the upper surface of the organic semiconductor layer through a physical absorption method and applying a bias voltage on the gate electrode. The present invention further discloses a DNA biosensor preparation method. According to the present invention, bias voltage applying is adopted to effectively improve DNA probe fixation and a hybridization efficiency of the DNA probe and DNA to be detected so as to increase sensor sensitivity; and compared with the conventional biosensor, the biosensor of the present invention has the following characteristics that: the same functions are achieved while a detection time is shortened.

The invention discloses a biological target molecule detection method based on multivalent capture and output signal amplification, and belongs to the technical field of biological detection. The detection method comprises the following steps: adding an identification probe into a solution to be detected, identifying and combining targeted biological target molecules, releasing an initiating sequence, allowing the initiating sequence to open hairpin probes to generate a hybridization chain reaction, adding an auxiliary probe, sealing hairpin probes which do not participate in the reaction, placing a carrier which is self-assembled with a capture probe on a gold interface in the reaction solution, capturing a hybridization chain reaction product onto a gold sheet, and finally analyzing thesignal mark change on the gold interface to further calculate the content of the targeted biological target molecules in the sample to be detected. The method forms a plurality of hybridization chainreaction products are formed through the hybridization chain reaction, and then utilizes interface multivalent capture and signal output amplification, so that the detection of target molecules is realized; and under the help of the auxiliary probe, the efficiency of capturing hybridization chain reaction products by the sensing interface is improved, and the detection sensitivity is improved.

The invention discloses a probe method detecting human TERT gene promoter mutation and a reagent kit thereof. The method includes the following steps that firstly, human genome DNA is added in a TERT C228T PCR reaction mixed solution so that a C228T reaction system can be formed and fluorescence quantitative PCR amplification can be performed, and / or human genome DNA is added in a TERT C250T PCR reaction mixed solution so that a C250T reaction system can be formed and fluorescence quantitative PCR amplification can be performed; secondly, a result is judged, wherein analysis is performed according to the mutation Ct value of PCR amplification. The reagent kit comprises the TERT C228T PCR reaction mixed solution and / or the TERT C250T PCR reaction mixed solution. The probe method and the reagent kit provide a high-speed, high-sensitivity and high-specificity TERT gene promoter mutation detection system clinically.

A novel nucleic acid probe for nucleic acid determination includes a single-stranded nucleic acid labeled with plural fluorescent dyes containing at least one pair of fluorescent dyes to induce FRET, the pair of fluorescent dyes including a fluorescent dye (a donor dye) capable of serving as a donor dye and a fluorescent dye (an acceptor dye) capable of serving as an acceptor dye, in which the nucleic acid probe has such a base sequence and is labeled with the fluorescent dyes so that the fluorescence intensity of the acceptor dye decreases upon hybridization with a target nucleic acid. A novel nucleic acid determination method uses the probe. The probe and method can determine one or more types of target nucleic acids in an assay system in parallel using a simple apparatus.

The invention discloses a preparation method for a long probe which can be used for multiplex ligation-dependent probe amplification (MLPA). The preparation method comprises the following steps: (1) selecting a vector which is irrelevant with a target sequence to be tested to serve as a template, and designing a primer according to the length of a target nucleotide sequence and the length of a target probe; (2) artificially synthesizing the primer; (3) performing polymerase chain reaction (PCR) amplification; (4) purifying and recovering PCR products; (5) digesting the PCR products by using Lambda exonuclease; and (6) recovering single-stranded deoxyribonucleic acid (ssDNA). The preparation method for the probe for the MLPA is low in cost, and all operations, such as the amplification, enzyme digestion and the like, can be finished only by a PCR instrument; recovery is performed by using a special ssDNA recovery kit after the enzyme digestion is finished, thereby, double-stranded DNA (dsDNA) and nucleotide which are not completely digested are eliminated, and the hybridization efficiency when an MLPA experiment is conducted is increased; and through full Lambda exonuclease digestion, the dsDNA can be changed into the ssDNA to the maximum degree, and thereby, the preparation efficiency of the probe is increased.

The invention discloses a method for separation, dispersion and sequence repetition of a bead-probe complex. The method comprises the following steps: firstly, the preparation of a genome fragment enriched library, which is to perform restriction enzyme digestion on a genome, to recover fragments with adequate dimensions, to add linkers on T4 ligase, and to perform cyclic amplification of the genome fragment library after restriction enzyme digestion through PCR of a small number of linkers; secondly, the preparation of the bead-probe complex, which is to realize biotin labeled DNA probes by the primer amplification method, and to bind biotin labeled single-chain probes on beads; and thirdly, acquisition of a target fragment group, which is to perform crossover on the bead-probe complex and the genome library, to acquire a target fragment, to separate and clone the target fragment, and to perform positive selection on the target fragment. The method is simple, high-efficiency and quick, shortens the experimental period, reduces the cost consumption, greatly improves the efficiency, and does not require special experimental equipment.

An automatic injection device comprises at least an injection unit (1). The said injection unit (1) is formed by sealing a cover plate layer (3) with hydrophilic surfaces and a microfluid layer (4). The said cover plate layer (3) is provided with at least two through holes (5). he said microfluid layer (4) is provided with a hollow-out hybridization chamber (7) and at least two hollow-out microfluid channels (6). One end of each channel (6) is connected with the hybridization chamber (7), and the other end is connected with a through hole (5) of the cover plate layer (3) respectively. Taking advantage of the hydrophilicity of the cover plate, the automatic injection device makes a solution automatically enter and fill the hybridization chamber (7) and the microfluid channels (6) by the driving force of liquid surface tension. The flow uniformity of sample solution in microarray chip is achieved by the structural design of the hybridization chamber (7) and the microfluid channels (6). The automatic injection device has advantages of simple manufacture, easy operation, high hybridization efficiency, low sample cost, and automatic quantificational injection.

The invention relates to the technical field of wheat hybridization, and discloses a distant hybridization method of wild oats and common wheat. According to the method, the common wheat is taken as a female parent, and the wild oats are taken as male parents; the female parent is planted in a dispersed way, the male parents are evenly planted around the female parent, and a one-female and multi-male hybridization system is formed by the female parent and the plurality of male parents planted around the female parent; or the male parent is planted in a dispersed way, a plurality of female parents are evenly planted around the male parent, and one-male and multiple-female hybridization system is formed by the male parent and the plurality of female parents planted around the male parent. After the distant hybridization method of the wild oats and the common wheat is adopted, the process of selecting male spike in an existing hybridization process is omitted, and the efficiency and the operation convenience of hybridization are greatly improved; furthermore, the male parents and the female parents are planted in a mixed way, so that the distant hybridization of the wild oats and the common wheat can be better realized, and the problem that the hybridization of the wild oats and the common wheat is not firm can be solved.

The invention provides a preparation method and application of a hydrogel electrode and relates to hydrogel electrodes. The preparation method of the hydrogel electrode comprises the following steps: with graphite powder as a raw material, adding sodium nitrate, sulphuric acid and potassium hypermanganate, mixing and then carrying out reaction until a thick mixture is formed; then adding pure water for the first time, further carrying out reaction, then adding pure water for the second time and stopping reaction, then adding a hydrogen peroxide solution for removing unreacted potassium hypermanganate, washing, centrifuging, and drying, so that graphite oxide solids are obtained, and carrying out ultrasonic treatment on the graphite oxide solids, so that a uniformly dispersed graphene oxide aqueous solution is obtained; mixing the graphene oxide aqueous solution with DNA of milt, then adding the mixture into a centrifugal tube to be heated, and after gel is stably formed, inserting a copper wire into a small hole in the bottom of the centrifugal tube to be fixed, so that a graphene oxide and milt-DNA compounded hydrogel electrode is obtained. The graphene oxide and milt-DNA compounded hydrogel electrode can be used for preparing a graphene oxide and milt-DNA composite hydrogel biosensor and can be applied to detection of mutation of mitochondrial DNA of ovarian cancer.

The invention provides a human papillomavirus gene chip, a preparation method and application thereof, an assay kit and application thereof. The gene chip comprises a solid-phase vector and an oligonucleotide probe fixed on the solid-phase vector, wherein the oligonucleotide probe contains a DNA fragment or a complementary DNA fragment thereof selected from L1 genes of the human papillomavirus or human beta globin gene. The invention also provides a preparation method for the gene chip and an assay kit containing the gene chip. The gene chip and the kit have the characteristics of simple and convenient operation, high flux, high accuracy, strong repeatability and the like, can achieve the aim of detecting 25 different types of human papillomavirus, and can be applied to the clinical test, epidemiology analysis and the like of medical and health departments.

The invention relates to examination mutant hepatitis virus method, especially the method of using DNA reverse dot blot hybridization technique to quickly and exactly distinguish clinic blood sample wild type and rummy fuding tolerance mutant hepatitis b virus. And it also relates to clinic measuring kit.

The invention relates to gene chip used for detecting functional central nervous damage diagnosis and the preparing method. said gene chip is equipped with oligonucleotide gene probe array, the oligonucleotide biomolecule of said probe array is the special biological gene factor playing improtant role in multiple central nervous system damage process simutaneously, relating to netural cell death and tissue necrosis, inflammation, irritated response, ion dysequilibrium inside cell, signal transmission, abnormal control of cell cycle, abnormal cell construction and nerve degeneration damage morbid physiology and molecular biology active gene, respectively. The gene chip can be used to check the conditioning way, expression level and change rule of biological factor relative to cerebral ischemia damage inside body in the beginning of clinic for high risk group of cerebral apoplexy, and to predict possibility of cerebral apoplexy; and used to conditioning way and change rule of special biological factor relative to nerve damage inside nerve cell after central nervous system damage, and relationship between cell death and disease occurancy, and provides checking criterion for cilinical treatment.

The invention discloses a method for selecting the hybrid seeds of Brassica juncea var. multiceps. Yongxue No.3 of Brassica juncea var. multiceps is bred through hybrid seed production of Ningbo fine-leaf yellow Brassica juncea var. multiceps cytoplasm male sterile line 07-50A and fine-leaf mustard 07-2 inbred line Brassica juncea var. multiceps as a female parent and a male parent respectively. The Ningbo fine-leaf yellow Brassica juncea var. multiceps cytoplasm male sterile line 07-50A is obtained by hybridizing a cytoplasm male sterile source as a female parent and Ningbo fine-leaf yellow Brassica juncea var. multiceps to obtain an F1 generation and carrying out backcross breeding by using the Ningbo fine-leaf yellow Brassica juncea var. multiceps as a recurrent parent; and the male parent fine-leaf mustard 07-2 inbred line Brassica juncea var. multiceps is obtained by the inbred purification of a local variety of fine-leaf mustard having excellent comprehensive properties. The new species Yongxue No.3 obtained through the method has the advantages of high output, strong disease resistances, high stalk-leaf ratio, excellent processing property, and suitableness for the plantation of winter Brassica juncea var. multiceps.

A method for improving peanut hybridization efficiency is characterized by comprising the steps of (1) removing main bolts when peanut maternal plant stems are 10 centimeters high, and spraying 1ml to 2ml of 4ppm 1.8% compound sodium nitrophenolate solutions to increase plant activity; (2) selecting strong maternal plants to prune flowers to prepare for hybridization; (3) detasseling alternative plants at 4 p.m. to 6 p.m., and performing pollination at 6 a.m. to 9 a.m. the next morning, and spraying mixtures of 100pp,m GA and 2.5% urea in a mass ratio of 1:1 in ten minutes after pollination; (4) removing all flowers on the maternal plants before 10 a.m. every day after all combined pollination is over; (5) smearing 100ppm IAA through cotton balls onto appearing peanut pegs after 4 days of smearing treatment; (6) smearing 50ppm6-BA on the marked peanut pegs after 12 days of smearing treatment; (7) spraying foliage fertilizers at the initial stage of podding to promote the growth of plants. The method has the advantages that the hybridization success rate is improved, false hybrids are removed, the plump rate and the two-peanut rate are improved, and pods are promoted to be ripe.

The invention belongs to the warm water male-killing technical field of japonica rice, and provides a method for killing male of japonica rice by warm water. The method comprises the steps of digging out to-be-hybridized female parents in the afternoon of the previous day of blooming; cutting too old and too tender tillers; removing bloomed glumous flowers and glumous flowers with tender base part; selecting glumous flowers which will bloom in 2-3 days, directly cutting 1 / 2 of the glumous flowers; in the morning of day for pollination, preparing one pot of warm water at 45 DEG C; soaking the cut japonica rice ears in the warm water at 45 DEG C in about 2 hours before the female parent blooms normally; after 3-5min, taking out the ears, and flicking extended anthers to enable stigma to be bared, wherein the anthers extend successively; covering the female parent with kraft paper bag after male-killing, transfering the female parent to place near male parent, and taking the male parent to pollinate when blooming; grouping the pollinated female parents for transplanting; cutting the hybrid seeds when maturing. The method for killing male of japonica rice by warm water breaks through hybridizing time limit of the japonica rice, increases hybridization efficiency, has good pollination effect, improves outcrossing maturing rate, is low in glumous flower-cutting technology demand and kills male thoroughly.

The invention relates to an RNA (ribonucleic acid) purification chromatin separation technique. The technique includes the following steps: binding probes with magnetic beads prior to hybridizing with chromatin lysis solution, and then performing washing for multiple times via gradient temperature. The magnetic beads are closed with BSA and random oligonucleotide primers in advance, and accordingly nonspecific binding of the magnetic beads with proteins and RNA is reduced. The magnetic beads are incubated with the probes firstly prior to being incubated with the proteins after excessive probes are removed, gradient temperature washing is performed after hybridization to assure that the magnetic beads fully bind with the probes and bind with target RNA, and hybridization efficiency is increased.

Methods, systems and compositions are provided for analyzing one or more nucleic acid molecules. The methods, systems and compositions may comprise one or more target specific-oligonucleotide probes (TSPs). The TSPs may hybridize to nucleic acid molecules that are less than or equal to 200 nucleotides in length. The nucleic acid molecules may be small RNA molecules (e.g., miRNA, ncRNA, siRNA, shRNA). The methods, systems and compositions fmd use in a number of applications, for example, isolation of nucleic acid molecules, analysis of low abundance nucleic acid molecules, and / or enrichment of nucleic acid molecules.

The invention relates to the field of hybridization breeding, and specifically relates to a soybean sexual hybridization method. The soybean sexual hybridization method comprises the following nine links: (1) fixing flower buds; (2) removing sprout for the first time, namely only reserving quasi-hybrid flower buds in the whole inflorescence, and completely removing the rest of flower buds; (3) removing calyx, namely removing calyx lobus at the upper half part of the calyx while reserving the basic cylindrical structure of the calyx; (4) castrating; (5) belaying, namely coiling a short wire into an annular slipknot to gently tie a flower stalk after passing through the flower; (6) taking flowers, namely before 6 a.m. the next day after castrating, picking flowers with unopened petals and uncracked carina on a plant with typical male parent variety characters as a pollen taking object; (7) pollinating at 6-8 a.m. the next day after castrating; (8) putting up brass plates; and (9) managing in the later period. The hybrid survival rate and hybrid efficiency are greatly improved according to the soybean sexual hybridization method provided by the invention.

A method of using Neutrilized DNA (N-DNA) as a surface probe for a high throughput detection platform is disclosed. FET and SPRi are used as high throughput detection platforms to demonstrate that the N-DNA surface probe produces good results and enhances detection sensitivity. The N-DNA modifies the charged oxygen ions (O−) on the phosphate backbone through methylation, ethylation, propylation, or alkylation, so that the backbone is not charged after this modification to increase the hybridization efficiency, sensitivity and to make the signal more clear.

The invention relates to a humidification temperature-controlled oscillation type hybridization device. The humidification temperature-controlled oscillation type hybridization device comprises a base, an upper cover, a lower heating plate and an upper heating plate, wherein the upper cover is matched with the base, the lower heating plate and the upper heating plate are respectively arranged on the base and the upper cover, and a heating cavity is formed by the upper cover and the base; and the humidification temperature-controlled oscillation type hybridization device is characterized in that the base is provided with a functional rack, the bottom part of the functional rack is connected with an oscillation motor and a controller which is in signal connection with the oscillation motor, the heating cavity is internally provided with a humidification device, the humidification device is in signal connection with the controller, the lower heating plate and the upper heating plate are in signal connection with a temperature control device, and the temperature control device is in signal connection with the controller. The humidification temperature-controlled oscillation type hybridization device disclosed by the invention has the beneficial effects that through the action of a humidification control device, the temperature control device and the oscillation motor, the purposes that the humidification can be continuously carried out within relatively-long time, the humidity can be regulated in real time according to test needs, and the oscillation can be simultaneously generated according to the test needs can be achieved by the humidification temperature-controlled oscillation type hybridization device, and the hybridization efficiency and the hybridization quality can be increased.

The invention discloses a gene chip for detecting transgenic cattle and a preparation method and application thereof. The chip provided by the invention contains solid phase carrier and an oligonucleotide probe fixed on the carrier, wherein the oligonucleotide probe contains a detection probe and a quality control probe; and the detection probe is a specific probe of mitochondrial genes of cattle, sheep and pig, human lactoferrin gene, human lysozyme gene, human-alpha-lactoalbumin gene, green fluorescent protein gene and neomycin phosphotransferase gene. By adopting the gene chip and the construction method to detect transgenic cattle, the accuracy, specificity and sensitivity are all higher; and the gene chip is suitable for the high throughout screening of transgenic cattle.

According to the present invention, there is provided a method for detecting a sequence of a nucleic acid in a sample which is complementary to a base sequence of a probe nucleic acid which comprises a step of hybridizing the nucleic acid in the sample with the probe nucleic acid, wherein one or more types of bases of the nucleic acid in the sample and / or the probe nucleic acid are chemically modified in advance. According to the method of the present invention, for example, viruses causative of diseases, exogenous genetic substances such as bacteria, or the presence, mutation or expression state of genes carrying particular genetic information of organisms including human, can be detected efficiently.