55results about How to "Short hybridization time" patented technology

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 discloses a kind of biological chip hybridisation machine, comprising hybridisation cavity, chip carrying disk, rotary part, temperature control part and master control part. The chip is installed in hybridisation cavity, chip carrying disk is drove by rotary part; rotary part is controlled by master part, the temperature inside hybridisation cavity is moderated by said temeprature control part. The invention aslo discloses an optimal method for controlling biological chip hybridisation process. It comprises following steps: (1) putting washing liquid into hybridisation cavity in advance, putting pre- hybrid biological chip box on chip carrying disk, rising temperature in hybridisation cavity through temperature part to proper temperature; (2) the rotary vibration of chip carrying disk is drove by controlling rotary part according to set vibration index, (3) drying left hybridisation liquid by rotary part after hybridisation is over.

The invention provides a nucleic acid hybridized platform based on micro-flow control and a hybridization analysis method thereof; the platform comprises an upper layer and a lower layer; the upper layer is a centrifugal micro-flow controlled chip, and the lower layer is a substrate of fixed oligonucleotide probe; the upper and the lower layers are sealed to form a fluid passage or a chamber; the hybridization analysis method takes a centrifugal force and a capillary tube force as driving forces to accelerate liquid flow to flow into a hybridized passage automatically in reciprocating manner; the chip is rotated to produce the centrifugal force, and the sample liquid flow collapses under the effect of the centrifugal force; the liquid flows through the hybridized passage and enters a temporary liquid collecting tank; then the chip is stopped rotating and the sample liquid flow is returned back to the hybridized passage from the temporary liquid collecting tank under the effect of the capillary tube force; and the repeated steps of rotating and stopping chip can reciprocate the liquid flow. The invention has the advantages of fast material transmission, short hybridizing time and low consumption of sample; and multiple samples can be analyzed in parallel simultaneously.

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.

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 provides a microfluidic-based nucleic acid hybridization reaction platform and a microfluidic-based nucleic acid hybridization analysis method. The platform consists of an upper layer and a lower layer, wherein the upper layer is a microfluidic chip controlled by a micropump and a microvalve; the lower layer is a substrate for fixing an oligonucleotide probe; and a fluid channel or a chamber is formed by the upper layer and the lower layer through irreversible sealing. The hybridization analysis method comprises a step of promoting liquid flow to automatically flow to and fro in a hybridization channel by taking positive pressure and negative pressure which are controlled by a program as a driving force, namely filling a sample into a microchannel by a capillary force or negative pressure after the sample enters a waste liquid chamber, and promoting liquid to flow to and fro in the microchannel by controlling the pushing and bounce (positive pressure and negative pressure) of two valve seats at both ends of the liquid channel. The microfluidic-based nucleic acid hybridization reaction platform and the microfluidic-based nucleic acid hybridization analysis method have the advantages that: mass transfer of materials is quick, hybridization time is short, the sample consumption is low, a hybridization signal of the sample can be detected in real time, the integration is easy to realize, a plurality of samples can be subjected to parallel analysis synchronously and the like.

The invention discloses a composite chip for rapid prenatal diagnosis of PKU and a method. The composite chip comprises a DNA chip and a micro-fluidic chip; the DNA chip is formed by fixing various point mutation PKU specific diagnostic gene probes on a glass chip; and the surface of the micro-fluidic chip provided with a passage is butted and automatically adsorbed firmly with the surface of the glass chip provided with the point mutation PKU specific diagnostic gene probes by taking a reaction area and a gene probe area as butting points. The method comprises the following steps: extracting DNA in plasma and leucocyte of a pregnant woman, performing amplification on exon genes of the DNA and performing reaction on a primer to obtain a sample; respectively adding the sample, washing fluid and a cleaning solution into a sample cell, a washing fluid cell and a cleaning solution cell of the composite chip; centrifuging the composite chip; and unveiling and discarding the micro-fluidic chip on the glass chip, and observing a result of the reaction of the sample and the reaction area. The composite chip and the method can realize the rapid prenatal diagnosis of the PKU.

The invention belongs to the field of biomedical clinical detection and relates to a kit for identification of circulating tumor cells through CD45 immunofluorescence-CEP17 fluorescence in-situ hybridization probe combined one-step method co-dyeing, and a use thereof. The kit comprises a fixing solution, 2*SSC, 0.3% NP-40 / 0.4*SSC, 0.075 M of a KCl solution, a CD45 monoclonal antibody-CEP17 probe mixed liquid, a DAPI redyeing agent, a perforating agent, a sealing liquid and a mounting medium. The kit uses combination of a CD45 immunofluorescent antibody and a CEP17 fluorescence in-situ hybridization probe to identify the captured circulating tumor cells, completes immunofluorescence and fluorescence in-situ hybridization detection by a one-step method, and solves the problem that the immunofluorescence detection and the fluorescence in-situ hybridization detection produce mutual interference in combined use and detection time is long. The one-step method-based hybridization incubation can be fast finished in 2h so that detection time is greatly reduced. The detection result is visual.

The invention discloses a method for rapid detection of BCR / ABL gene fusion by using fluorescence in-situ hybridization technology. The method is characterized in that detection of each target site needs three kinds of probes, i.e., contact probes, amplification probes and fluorescence probes; the 5' terminals of the contact probes specifically bind to a BCR gene (or an ABL gene), and the 3' terminals of the contact probes both have 20 sets of a repetitive sequence consisting of 20 bases and can specifically bind to the amplification probes; the 5' terminals of the amplification probes specifically bind to the 3' terminals of the contact probes, and the 3' terminals of the amplification probes both have 20 sets of a repetitive sequence consisting of 20 bases and can specifically bind to the fluorescence probes; the 5' terminals of the fluorescence probes are labeled by CY3 or FITC; and through synergism of the three kinds of probes, a fluorescence signal is amplified by 400 times, and thus, rapid and sensitive detection of BCR / ABL gene fusion is realized. The invention also discloses a kit and probe set for rapid detection of BCR / ABL gene fusion.

A method of detecting or analyzing a target sequence in a genomic DNA by using a capture probe immobilized on a solid carrier includes: bringing the target nucleic acid into contact with a first query probe that has a sequence complementary to a portion of the target sequence or to a sequence adjacent to the portion and a second query probe that has a sequence complementary to another portion of the target sequence or to a sequence adjacent to the another portion and that has a recognition sequence complementary to a portion of the capture probe; acquiring a ligated molecule by ligating the first query probe and the second query probe that are hybridized to the target nucleic acid; bringing the ligated molecule into contact with the capture probe on the solid carrier and then capturing the ligated molecule on the solid carrier by hybridizing the capture probe with the recognition sequence in the ligated molecule; and detecting the captured ligated molecule.

The invention discloses a genome sequencing library hybrid capture kit, a washing kit and applications of the kits in liquid-phase hybrid capture. The capture kit consists of a 2*concentration solution and 100% de-ionized formamide, wherein the 2*concentration solution comprises 100-150mM of sodium dihydrogen phosphate, a 4*sodium citrate buffer solution, 1-3mM of ethylene diamine tetraacetic acid, a 2*Denhardt solution, 2-5mg / ml of Carrier RNA and 15-20w / w% of dextran sulfate, wherein the 4*sodium citrate solution comprises 0.6M of sodium chloride and 0.06M of sodium citrate and pH value of the 4*sodium citrate solution is 7.0; and the 2*Denhardt solution comprises 0.04% of polysucrose 400, 0.04% of polyvinylpyrrolidone and 0.04% of bovine serum albumin.

The invention discloses a PML-RAR alpha fusion gene detection kit and a detection method thereof. The kit comprises a first group of probes for detecting an upstream gene sequence of an L-type breakage hot spot of a PML gene and a second group of probes for detecting all gene sequences of a RAR alpha gene. Each one of probes is labeled a dye. The dyes labeling the two groups of probes are in different colors. The two groups of probes are amplification products obtained by amplification based on a human epigenome DNA as a template and amplification primers. The probes have strong specificity and low background noise and realizes the optimal balance between the detection specificity and hybridization time. The PML-RAR alpha fusion gene detection kit guarantees result specificity and sensitivity, shortens hybridization time, has detection time of 7-8h and improves detection efficiency.

The quick detection method of pathogenic microbe diagnosis type gene chip includes the following steps: preparing slide; preparing probe of length from several hundred to several thousand base and concentration of 0.25-0.5 microgram / microliter via PCR proliferation of genes and specific segments in pathogen genome; preparing chip via sample application with gene chip sample applying instrument; printing and processing; processing sample via incorporating marker Cy5-dUTP in PCR proliferation and introducing restrictive enzyme incision to obtain segment of 200-600 bps length; hybridizing in 2X hybridizing liquid comprising 60 % formamide, 12XSSC and 0.24 % SDS; cleaning after hybridization; and scanning detection. The present invention has the advantages of high marking efficiency, stable hybridizing detection result, greatly shortened hybridization time, etc. and is especially suitable for the diagnosis of infectious diseases clinically.

The invention relates to a solid phase DNA chip hybridization solution and a hybridization method. The solid phase DNA chip hybridization solution is composed of raw materials of NaCl, Na-MES, EDTA and SDS, wherein NaCl is 717mm, Na-MES is 400mm, EDTA is 100 mu m, and SDS accounts for 0.2%(w / t). The main ingredients of the hybridization solution are as follows: NaCl, Na-MES, EDTA and SDS. During hybridization, a marked PCR product is only dried and is mixed and dissolved by preheated hybridization solution and deionized water at the ratio of 1:1, and the obtained mixture is added into an incubation chamber to be incubated. The method of the invention is simple and easy to realize, can improve hybridization temperature and can effectively shorten hybridization time. The hybridization solution can preferably control a background value generated by hybridization, improves hybridization signal, reduces interference of the background value to the signal value and optimizes the hybridization effect.

The invention relates to a primer probe group, a kit and a detection method for PCR detection of a potato broom top virus, and belongs to the technical field of molecular biology. In order to solve the problem that the PMTV cannot be accurately and rapidly detected by an existing detection method, the invention provides a primer probe group for PCR detection of the potato broom top virus. The primer probe group comprises a primer probe group-PMTV2 and a primer probe group-PMTV3; the invention also provides a detection kit comprising the primer probe group and a detection method. According to the invention, cDNA obtained through RNA reverse transcription of a sample to be detected is taken as a template, and primers in a primer probe group are adopted to carry out duplex PCR reaction; the obtained PCR reaction product is hybridized with a chip connected with the probes in the primer probe group, developing is conducted, and if the corresponding position of the chip is bluish violet, it is judged that the chip is positive. According to the invention, the defect that naked virus RNA cannot be detected by ELISA is avoided, and false positive and false negative conditions of an RT-PCR method are reduced.

The invention discloses a kit and a detection method for abnormity of ROS1 and C-met genes. The kit includes a first group of probes and a second group of probes, which aim to the ROS1 gene, and / or a third group of probes and a fourth group of probes, which aim to the C-met gene. Each group of probes is labeled with a fluorescent dye, wherein the color of the fluorescent dye on the probes in the same group is same while the colors of the fluorescent dyes on the probes in different groups are different. The four groups of probes respectively are amplification products by amplifying a primer with corresponding BAC cloning as a template. The FISH probes are produced through repeated comparison and selection and then utilization of optimized BAC cloning as the template and through amplification by designing a primer aiming to a non-repeated and highly-conserved sequence, so that the kit has excellent specificity and lower background noise. The kit can achieve the optimal balance between detection specificity and hybridization time length, so that not only are specificity and sensitivity of a result ensured, but also hybridization time is reduced, thereby increasing detection efficiency.

The invention relates to an AMPK activator (such as for instance metformin) for use in preventing metastasis in a patient suffering from a cancer, wherein said patient has a non-mutated p53 gene or lacks a mutant form of the p53 protein. The invention also relates to an AMPK activator for use in improving the survival time of a patient suffering from a cancer, wherein said patient has a non-mutated p53 gene or lacks a mutant form of the p53 protein.

The invention discloses an Oligo probe and a preparation method and application thereof. The preparation method comprises: mixing a first partial sequence and a second partial sequence, carrying out denaturation and annealing in a labeled deoxyribonucleotide system, and adding a DNA polymerase into the system for a polymerization reaction to obtain the Oligo probe. The first partial sequence is acommon sequence 1 connected to the end 3' of a specific oligonucleotide sequence and the second partial sequence is a common sequence 2 connected to the end 3' of a labeled sequence. The specific oligonucleotide sequence is specifically bonded to a gene to be detected and does not contain the repeated sequence. The common sequence 2 is a sequence with a modified end 3' and the common sequence 1 iscomplementary with the common sequence 2. The Oligo probe has high sensitivity and specificity and can be used for preparation of a FISH probe. The FISH probe easily enters cells and has a fast hybridization reaction rate. Through a specific hybridization buffer, the hybridization time can be shortened to 1 to 2h.

The invention discloses a primer pair, a probe and a kit for detecting SLCO1B1 521T)C gene polymorphism. The kit comprises a primer and a fluorescence probe; the primer is designed for SLCO1B1 521T locus and has a base sequence shown as SEQ ID No.1 and SEQ ID No.2, wherein a 5'-base of the base sequence is equipped with an amino-modified group; the fluorescence probe is designed for SLCO1B1 521T locus and has the base sequence shown as SEQ ID No.3 and SEQ ID No.4, wherein one basic group in the base sequence is equipped with the amino-modified group. Compared with the prior art, the kit disclosed by the invention adopts the amino modified primer and probe, further limits the detection time and is capable of obviously shortening the time required by detection, reducing non-specific amplification, detecting SNPs and point mutation, reducing design difficulty and detection cost without influence on diagnosis accuracy, realizing a method for detecting mutation without opening a single reaction tube, extremely avoiding aerosol pollution, bringing convenience to clinic application and reducing the error caused by operation.

The invention belongs to the field of molecular biology, in particular to a fluorescence in situ hybridization probe for detecting KIAA1549-BRAF fusion gene and preparation method and application thereof. The fluorescence in situ hybridization probe comprises two probe libraries, a KIAA1549 gene hybridization probe library: the starting position of the probe is designed in the range of 200 Kb around the KIAA1549 gene breakpoint, the probe extends toward the direction of the centromere, and the length of the probe is 300 to 1000 Kb; BRAF gene hybridization probe library: The starting position of the probe is designed in the range of 200 Kb around the breakpoint of BRAF gene, and the probe extends to the telomere. The length of the probe is 300 - 1000 Kb. The invention designs probes in twofusion gene outer regions respectively, realizes the detection of two fusion genes which are relatively close on the same chromosome, and can directly reflect the fusion state of the two genes on thecell level.

The invention discloses a primer pair, a probe and a kit used for detecting polymorphism of an MTHFR gene. The kit comprises: the primer with the base sequences represented by SEQ ID No.1 and SEQ ID No.2, which is designed for the c.677 site of the MTHFR gene, wherein the first base at the 5' end of the base sequence represented by the SEQ ID No.1 has an amino group modification group, and the first base at the 5' end of the base sequence represented by the SEQ ID No.2 has an amino group modification group; and the probe with the base sequences represented by SEQ ID No.3 and SEQ ID No.4, which is designed for the c.677 site of the MTHFR gene, wherein one base of the base sequence represented by the SEQ ID No.3 has an amino group modification group, and one base of the base sequence represented by the SEQ ID No.4 has an amino group modification group. Compared with kits in the prior art, the kit disclosed in the invention has the advantages of remarkable shortening of the detection time, reduction of nonspecific amplification, detection of SNPs and point mutation, reduction of the design difficulty and the detection cost, no influences on the diagnosis precision, realization of mutation detection using a single reaction tube without cover opening, maximal avoiding of aerosol pollution, convenience in clinic use, and reduction of operation induced errors.

The invention discloses a kit for detecting abnormity of BRCA1 and PTEN genes and detection method. The kit includes a first group of probes and a second group of probes, which aim to the BRCA1 gene, and / or a third group of probes and a fourth group of probes, which aim to the PTEN gene. Each group of probes is labeled with a fluorescent dye, wherein the color of the fluorescent dye on the probes in the same group is same while the color of the fluorescent dye on the probes in different groups are different. The four groups of probes respectively are amplification products by amplifying a primer with corresponding BAC cloning as a template. The FISH probes are produced through repeated comparison and selection and then utilization of optimized BAC cloning as the template and through amplification by designing a primer aiming to a non-repeated and highly-conserved sequence, so that the kit has excellent specificity and lower background noise. The kit can achieve the optimal balance between detection specificity and hybridization time length, so that not only are specificity and sensitivity of a result ensured, but also hybridization time is reduced, thereby increasing detection efficiency.

The invention discloses a probe group for detecting NTRK fusion mutation. The probe group comprises one or more probes of which the nucleotide sequences are shown as SeqID NO. 1-146. The invention also discloses a kit for detecting NTRK fusion mutation. The kit comprises the probe group. The invention further provides a method for detecting the NTRK fusion mutation by using the probe group. The method for detecting the NTRK fusion mutation by using the probe group designed by the invention shortens the hybridization time, and can detect whether the NTRK gene of a non-small cell lung cancer patient is subjected to fusion mutation or not more quickly. Compared with a detection gold standard FISH method, the method has the advantages of high detection sensitivity, wide coverage, high flux andthe like, and is beneficial to large-scale clinical development.

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). The 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 a gene chip for enrichment and extraction of transgenic components and a preparation method of the gene chip. The preparation method comprises the following steps: establishing a transgenic information database including the target gene sequence information and the general element information; then preparing a transgenic component capture probe; designing a connecting armmolecule and connecting the connecting arm molecule with the 5' end of the probe molecule; processing the surface of a solid-phase substrate to prepare an electrode substrate; and fixing the transgenic component capture probe on the surface of the solid-phase substrate through the connecting molecule at the tail end of the connecting arm, so as to obtain the gene chip for enrichment and extractionof the transgenic components. According to the invention, the hybridization efficiency and the hybridization speed of the transgenic components with the extraction chip probe molecule are improved byadopting the combination of the connecting arm and electric field acceleration, and the hybridization time is obviously shortened; and subsequently, the transgenic components bounded by hybridizationare separated from the capture probe through DNA degeneration and rapidly depart from the surface of the extraction chip under the action of a reverse electric field, so that the working efficiency of the chip is obviously improved.

The invention belongs to the field of biochemistry, in particular to the field of measurement or detection related to biochemistry. More particularly, the invention relates to a method for preparing aFISH probe of an EGFR gene, and a probe prepared by the method. The method of the invention uses BAC cloning, enzyme digestion interruption and asymmetric amplification to prepare a probe library, thereby effectively improving product sensitivity, specificity and signal strength.

The invention belongs to the field of biochemistry, in particular to the field of measurement or detection related to biochemistry. More particularly, the invention relates to a method for preparing aFISH probe of an EGFR gene, and a probe prepared by the method. The method of the invention uses BAC cloning, enzyme digestion interruption and asymmetric amplification to prepare a probe library, thereby effectively improving product sensitivity, specificity and signal strength.

The invention discloses an Oligo probe as well as its preparation method and application. In the present invention, the Oligo probe is obtained by mixing the first part sequence and the second part sequence, denaturing and annealing in a system with labeled deoxyribonucleotides, and then adding DNA polymerase for polymerization reaction; wherein, the first part The sequence is formed by connecting the common sequence 1 at the 3' end of the specific oligonucleotide sequence; the second part of the sequence is formed by connecting the common sequence 2 at the 3' end of the marker sequence; the specific oligonucleotide sequence is specific to the gene to be tested A sequence that combines and removes repetitive sequences, the consensus sequence 2 is a sequence modified at the 3' end, and the consensus sequence 1 and the consensus sequence 2 are complementary. The Oligo probe has high sensitivity and specificity, and can be used to prepare FISH probes. The obtained FISH probes are easy to enter cells, and the hybridization reaction is fast. Using a specific hybridization buffer, the hybridization time can be shortened to 1‑2h.

Provided is a novel approach for generating oligonucleotide probes and the use of these probes in gene expression profiling, by hybridization to test oligonucleotides on arrays or beads. This approach involves labeling of the complement oligonucleotide probes using a mixture of dye or hapten labeled-ddNTPs in solution. The labeled oligonucleotide probes are then used to hybridize to the test oligonucleotides on the solid support. Success in hybridization is monitored by associated signal on the solid support. This approach greatly reduces hybridization time, due to the simplification of the probe content. It is especially useful when analyzing a small number of genes, such as a signature set of genes for a disease or condition.

The invention provides a probe and a kit for detecting a Top2A gene as well as a detection method. The sequence of the probe provided by the invention is designed by aiming at a genome non-repetitivearea, so non-specific reaction is reduced and background signals are reduced; the probe provided by the invention is single-chain DNA, so the own pairing of the probe is reduced and the hybridizationefficiency is improved; and the probe provided by the invention is small in segment, can be rapidly combined with a targeted sequence and shortens the hybridization time.