114 results about "Nucleic Acid Probes" patented technology

The embodiment of the invention provides a connecting unit for chemical modification of a nucleic acid end group, a preparation method of the connecting unit and a nucleic acid probe. The connecting unit for chemical modification of the nucleic acid end group has a triethylene glycol structure, and the connecting unit has the following structure. Therefore, the connecting unit with the triethylene glycol structure is relatively simple in synthesis process, relatively high in yield, efficient in synthesis, good in nucleic acid end group chemical modification effect, applicable to connection of various types of probe molecules and nucleic acid molecules, capable of simply and efficiently preparing nucleic acid probes and wide in application range.

The invention provides a primer / probe composition for capillary electrophoresis analysis, a detection method and application. The primer / probe composition comprises a forward primer, a reverse primer and a nucleic acid probe, the 5'end of the nucleic acid probe carries a fluorescent label, and both the forward primer and the reverse primer do not carry the fluorescent label. Different targets are identified by detecting the nucleic acid probe, so that the length of an amplification product is not limited, only the nucleic acid probe carries a fluorescence signal without generating a non-specific fluorescence signal, the generation of a supersaturated fluorescence signal can be effectively prevented by using the nucleic acid probe with a specific concentration, the fluorescence signals among different targets do not interfere with each other, and the detection sensitivity is high. And multicolor fluorescence can be used at the same time.

The invention provides a nucleic acid probe, a primer group, a kit, a detection method and application, and relates to the technical field of biology. The nucleotide sequence of the nucleic acid probe is as shown in SEQ ID NO. 1; wherein the 3'end of the nucleic acid probe is modified with MGB, and locked nucleic acid is modified at a base position corresponding to the difference between the swine fever wild strain and the swine fever vaccine strain in the nucleotide sequence. According to the nucleic acid probe, a specific sequence is utilized to precisely target a swine fever wild strain, and locked nucleic acid modification is introduced at a differential site, so that the recognition capability on single-base mismatch is remarkably enhanced, non-specific binding to a vaccine strain is thoroughly blocked, and false positive interference is eliminated; and 3 '-terminal MGB modification is matched to improve the binding stability and sensitivity, so that the efficient and accurate identification of the swine fever wild strain under the immune background is ensured.

The present invention generally relates to systems and methods for imaging or determining nucleic acids, for instance, within cells. In some embodiments, the transcriptome of a cell may be determined. Certain embodiments are directed to determining nucleic acids, such as mRNA, within cells at relatively high resolutions. In some embodiments, a plurality of nucleic acid probes may be applied to a sample, and their binding within the sample determined, e.g., using fluorescence, to determine locations of the nucleic acid probes within the sample. In some embodiments, codewords may be based on the binding of the plurality of nucleic acid probes, and in some cases, the codewords may define an error-correcting code to reduce or prevent misidentification of the nucleic acids. In certain cases, a relatively large number of different targets may be identified using a relatively small number of labels, e.g., by using various combinatorial approaches.

The present invention generally relates to systems and methods for imaging or determining nucleic acids in cells or other samples. In some cases, the transcriptome of a cell may be determined. Certain embodiments are generally directed to determining nucleic acids and other targets in a sample at relatively high resolutions. For instance, nucleic acid probes may be applied to sample, and binding of the nucleic acid probes to a target may be amplified using primary and secondary amplifier nucleic acids. In some cases, there is a maximum number of amplifier nucleic acids that can be bound to a target, e.g., the binding is saturatable, and cannot grow indefinitely, even in the presence of abundant reagents. This may be advantageous, for example, for controlling the brightness of each binding event, controlling the size of the amplified regions (e.g., during imaging), and / or for limiting the degree of amplification noise (i.e. the final variation in amplified signal from molecule to molecule), etc. In addition, in some embodiments, the primary and / or secondary amplifier nucleic acids may be formed from only 3 of the 4 naturally-occurring nucleotides, which may result in less secondary structure, faster binding rates, etc. These properties can in some cases facilitate the rapid design of multiple orthogonal amplification sequences, allowing the extension of such an approach to many distinct molecular targets.

The invention discloses a multi-nucleic acid probe rapid modification and multi-target detection device based on electric field regulation and control. An instrument main body in the device is integrated with an automatic liquid path system and a micro electrochemical workstation module, and is used for realizing programmable reagent and sample conveying and multi-channel electrochemical measurement. The detachable sensing chip is formed by bonding a microelectrode array chip and a micro-fluidic chip, and is switched through a PCB (Printed Circuit Board) to realize plug and play. According to the present invention, the positive suction-negative discharge strategy of the electric field is utilized, the preferred positive potential is applied to the target electrode to efficiently attract the probe, and the preferred negative potential is applied to the non-target electrode to inhibit the non-specific adsorption so as to achieve the spatial selective rapid fixation and the accurate density regulation and control of the multi-nucleic acid probe in the extremely short time; the device can be used for quantitatively detecting various body fluid disease markers in a plurality of urine samples at the same time in single operation, and an efficient, accurate and portable multi-marker comprehensive analysis solution is provided for early screening and instant diagnosis of tumors.

The present invention generally relates to systems and methods for imaging or determining nucleic acids, for instance, within cells. In some embodiments, the transcriptome of a cell may be determined. Certain embodiments are directed to determining nucleic acids, such as mRNA, within cells at relatively high resolutions. In some embodiments, a plurality of nucleic acid probes may be applied to a sample, and their binding within the sample determined, e.g., using fluorescence, to determine locations of the nucleic acid probes within the sample. In some embodiments, codewords may be based on the binding of the plurality of nucleic acid probes, and in some cases, the codewords may define an error-correcting code to reduce or prevent misidentification of the nucleic acids. In certain cases, a relatively large number of different targets may be identified using a relatively small number of labels, e.g., by using various combinatorial approaches.

The invention relates to the technical field of molecular diagnosis and graphic processing, in particular to an electronic microarray chip dot matrix negative and positive judgment method based on an ISFET sensor. A chip hole site corresponds to an ISFET sensor, and after a hole bottom specific nucleic acid probe and a target sequence are hybridized and extended, the voltage of the sensor is changed. The method specifically comprises the following steps: data preprocessing: filtering median values of multi-frame electric signals of a hole site and taking a mean value; the dot matrix identification comprises the steps of threshold demarcation, Canny edge detection, circular Hough transform, optimal circle screening by Kdtree, array numbering and abnormal deletion processing to position a dot matrix; and calculating a result, extracting a signal difference value of the D1 stage and the D4 stage, evaluating numerical values of the positive quality control array and the to-be-detected sample array through a numerical value of the negative quality control array, and judging a final calculation result. According to the method, response changes are rapidly analyzed through image recognition, false detection and missing detection caused by numerical value differences in different arrays are reduced through multiple times of detection, and the detection accuracy is improved by selecting the most possible area through tickets.

The present disclosure is generally directed to anchoring bridge nucleic acid probes in a polymer matrix, which are configured to hybridize to cellular nucleic acid, embedding in a polymer matrix, clearing cellular components including the cellular nucleic acid, contacting the anchored bridge probes with primary probes that hybridize with the bridge probes and imaging. The disclosure describes methods for imaging cellular RNA in an RNase insensitive method.

The invention discloses a preparation method of a space chip, the space chip and application of the space chip, the preparation method comprises the following steps: hybridizing: connecting a single-stranded nucleic acid molecule to the surface of a solid-phase substrate, the single-stranded nucleic acid molecule comprises a first fixed sequence, a space bar code sequence, a sequencing primer binding sequence and a second fixed sequence, the surface of the solid-phase substrate of the chip substrate is connected with a first probe, and the first probe comprises a first fixed sequence complementary sequence and a second fixed sequence complementary sequence; carrying out ring closing to enable the single-stranded nucleic acid molecules to form annular nucleic acid molecules; carrying out rolling circle amplification; sequencing to obtain sequence information of the space bar code sequence; synthesizing single-stranded nucleic acid molecules by taking the rolling circle amplification product as a template to obtain a single-stranded nucleic acid molecule library; fixing and clustering: fixing the single-stranded nucleic acid molecule library on the surface of a solid-phase substrate to form a single-stranded nucleic acid molecule cluster. According to the invention, the chip with the nucleic acid probe carrying spatial position coordinate information can be prepared.

The present disclosure relates to finely modulated ultra-specific nucleic acid hybridization probes. The present invention provides compositions and methods for highly specific nucleic acid probes and primers. The probe system includes a complementary strand and a protective strand to partially form a double-stranded probe. The reaction standard free energy ([delta] G DEG rxn = [delta] G DEG t-TC-[delta] G DEG nh-PC + ([delta] G DEG v-TC-[delta] G DEG h-PC)) of hybridization between the probe and the target nucleic acid determined by formula 1 is about-4 kcal / mol to + 4 kcal / mol. Alternatively, the reaction standard free energy of hybridization between the probe and the target nucleic acid determined by Formula 1 is within 5 kcal / mol of the standard free energy determined by Formula 2 (-R [tau] ln (([P] 0-[C] 0) / [C] 0)), the term [P] 0 of Formula 2 being equal to the concentration of the protective strand, and the term [C] 0 of Formula 2 being equal to the concentration of the complementary strand. In addition, the present invention provides a method for the immediate fine modulation of a reaction using the probe of the present invention.

The invention discloses a human papilloma virus mRNA (messenger ribonucleic acid) detection kit for auxiliary diagnosis of cervical cancer patients. The human papilloma virus mRNA detection kit comprises a nucleic acid probe group and a series of nucleic acid probes AMP1-AMP5, wherein the nucleic acid probe group is used for being combined with a human papilloma virus mRNA target, and the nucleic acid probes AMP1-AMP5 are used for signal amplification. The human papilloma virus mRNA detection kit provided by the invention can assist pathologists in identifying and diagnosing cervical cancer patients, and provides a basis for clinicians to formulate a therapeutic schedule.

The invention discloses a synthesis method of 2 '-O-methyladenosine, and belongs to the technical field of nucleoside synthesis and medicinal chemistry. According to the method, xylose which is rich in source is taken as an initial raw material, eight-step reaction including acetalation protection, acetylation, glycosylation, deacetylation, methylation, deacetalation protection, Mitsunobu reaction and secondary deacetylation is performed, reaction conditions of each step are accurately controlled, and finally, a target product is synthesized. The method has the advantages of high methylation reaction regioselectivity, mild reaction conditions, cheap and easily available raw materials and controllable cost, and overcomes the problems of poor methylation reaction regioselectivity, harsh reaction conditions and high cost in the existing synthesis method. The obtained product has important application value in the fields of research and development of antiviral drugs, design of nucleic acid probes and the like.

An L-form ribonucleic acid (L-RNA) aptamer-antisense oligonucleotide (ASO) conjugate comprise the L-RNA aptamer, which comprises a ribonucleic acid sequence. The ASO comprises a deoxyribonucleic acid sequence selected from a group. A method of imaging amyloid precursor protein (APP) rG4 in a cell comprising transfecting the cell with a messenger RNA of APP, permeating the cell, contacting the permeated cell with a cyanine3 (Cy3) labeled APP nucleic acid probe and the L-RNA aptamer-ASO conjugate, and subjecting the product to fluorescence microscopy analysis to produce an image of the APP rG4 in the cell.

The invention discloses an exosome detection method based on double-hairpin nucleic acid probe assembly, and belongs to the technical field of exosome detection. The exosome detection method comprises the following steps: step 1, constructing a multi-element hairpin probe H1; step 2, constructing a multi-element hairpin probe H2; 3, preparing a probe; 4, multi-element signal probes are assembled, in the step 4, three groups of H1 probes and H2 probes are arranged in total, the proportion of the first group of H1 probes and 400 nM H2 probes is 80 nM, the proportion of the second group of H1 probes and 300 nM H2 probes is 80 nM, and the proportion of the third group of H1 probes and 350 nM H2 probes is 80 nM; according to the exosome detection method based on double-hairpin nucleic acid probe assembly, parallel detection of different exosome surface markers can be achieved by constructing multiple hairpin probes H1 and H2, and meanwhile the requirements for high specificity and stability are met.

The present disclosure relates in some aspects to methods and systems for analyzing a biological sample comprising contacting the biological sample with a nucleic acid probe non-covalently associated with a monomethine cyanine dye or salt thereof. In some aspects, a signal associated with the monomethine cyanine dye or salt thereof is detected to determine the location of a target analyte in the biological sample.

To provide an improved probe achieving enhanced brightness and producing a further lowered signal-to-noise ratio.SOLUTION: A compound useful as a nucleic acid probe is disclosed. In some embodiments, the compound has the following structure (I), or is a stereoisomer, tautomer, or salt thereof. Methods relating to preparation and use of such a compound are also provided.SELECTED DRAWING: None

The invention discloses a biochemical sensing system based on a catalytic hairpin assembly circular reaction and a G quadruplex-heme DNA enzyme formation reaction. The biochemical sensing system is used for photoelectrochemical colorimetric detection of circular RNA (circRNA). After the nucleic acid probe H1 is combined with the nano magnetic beads (MNPs), the circRNA is specifically recognized, and an MNPs / H1 / circRNA compound is formed; the MNPs / H1 / H2 compound reacts with a probe H2 to form an MNPs / H1 / H2 compound; the H1 probe and the H2 probe release a part of sequences by utilizing restriction endonuclease cutting, and the unreleased sequences are combined with hemin to form G quadruplex-heme DNA enzyme with peroxidase-like activity; through cascade catalytic reaction, the color of a solution of a reaction system is changed, the linear relation between the absorbance of the solution and the concentration of circRNA in the solution is fitted, and an effective method for carrying out photoelectrochemical colorimetric detection on circRNA is developed.

The invention belongs to the technical field of small molecule detection, and relates to a nucleic acid-based algal toxin on-site rapid quantitative detection paper chip device and a detection method thereof. A reaction area of the paper chip device is coated with an aptamer c-DNA and a fluorescent group labeled-DNA, and a reading area of the paper chip device is coated with a capture nucleic acid probe d-DNA; the nucleic acid sequence of the aptamer c-DNA is as shown in SEQ ID NO.1, and the 5'end of the c-DNA is marked with biotin; the nucleic acid sequence of the capture nucleic acid probe is shown as SEQ ID NO.1, the nucleic acid sequence of the capture nucleic acid probe is shown as SEQ ID NO.2, the 5'end of the capture nucleic acid probe is marked with an FAM fluorophore, the nucleic acid sequence of the capture nucleic acid probe d-DNA is shown as SEQ ID NO.3, and the 5 'end of the capture nucleic acid probe d-DNA is marked with biotin. The whole detection process does not exceed 10 minutes, the cost is low, and rapid on-site quantitative detection of algal toxin in water is achieved.

The application belongs to the technical field of molecular detection, and discloses a medicinal material identification system and method; the method comprises the following steps: emitting a topological protection acoustic wave to concentrate DNA of medicinal materials, marking the DNA with a CdTe quantum dot-nucleic acid probe, scraping the sample to extract DNA and performing PCR amplification, designing a logic gate staple chain to prepare a light-controlled origami probe, and injecting a PZT chip nano-pore array to be light-activated, and then, through AFM analysis of free energy, the medicinal materials are identified; the application constructs a complete technical chain from molecular recognition to reliable storage, and comprehensively solves multiple technical bottlenecks of traditional methods in medicinal material identification.

The application discloses a circRNA detection method based on Argonaute protein and rolling circle amplification. The method comprises the following steps: mixing a sample containing target circRNA to be detected with a specific reverse transcription primer and reverse transcriptase to obtain long-chain cDNA; mixing the obtained long-chain cDNA with pfAgo, gDNA and a DNA nucleic acid probe to carry out a reaction, and calculating the content of the target circRNA according to the fluorescence signal of the reaction mixture; the nucleotide sequence of the gDNA is specifically complementary to the cDNA sequence obtained by reverse transcription of the circRNA; the sequence of the DNA nucleic acid probe is specifically complementary to the sequence of the cDNA after one cutting of the cDNA by the pfAgo; and the DNA nucleic acid probe is a single-stranded DNA probe with a fluorescent label. The application can realize high-sensitivity, high-specificity and multiple target simultaneous analysis and detection of potential diagnostic biomarkers of liver cancer such as hsa_circ_0001445 and hsa_circ_0001141.

The application discloses a kind of photoactivation manganese ion bridging type deoxyribozyme probe preparation and its application in Cistanche inhibiting acetylcholinesterase activity.The probe is pSD@MnO2 Composite structure, which is composed of manganese dioxide nanosheet and functional nucleic acid assembly adsorbed on its surface, the assembly includes S chain labeled with fluorescence / quenching group and containing deoxyribozyme cleavage site, and pD chain partially hybridized with S chain and containing photocleavage site.Its preparation includes nucleic acid probe assembly, MnO2 Nanosheet complexing and purification steps.When applied, AChE catalytic product decomposes MnO2, releases Mn 2+ And nucleic acid chain; after centrifugation, pD chain is cut by ultraviolet light activation trigger, and released DNAzyme is cut under the assistance of Mn 2+ It produces amplified fluorescence signal; by comparing signal change before and after adding inhibitor, inhibition activity can be quantitatively evaluated.The application combines photocontrol activation with enzyme cascade amplification, realizes high sensitivity and operation synchronism of detection process, and is especially suitable for high-throughput rapid screening of AChE inhibitor in Cistanche and other natural products.

The present disclosure provides a chitin film-based biosensor, a preparation method and a kit. Specifically, the biosensor comprises a chitin film layer, a first sensing area and a second sensing area; an attachment film layer arranged in the first sensing area; a wire layer arranged in the second sensing area; an electrochemical sensing layer arranged in the second sensing area and in communication with the wire layer; and a nucleic acid probe set, wherein the nucleic acid probe set comprises a first nucleic acid sequence, a second nucleic acid sequence, a third nucleic acid sequence and a fourth nucleic acid sequence. By simultaneously arranging the first sensing area and the second sensing area on the chitin film layer, and by means of the nucleic acid probe set, the fluorescence detection of the analyte in the first sensing area and the electrochemical detection of the analyte in the second sensing area, the high-sensitivity and multi-modal detection of the analyte is realized, and the biosensor has a wide application prospect in the field of biological detection.

The application discloses a functional nucleic acid probe and a preparation method and application thereof. The preparation method comprises the following steps: forming a DNA sheet by self-assembly of a first DNA single strand and a second DNA single strand, and then combining the DNA sheet with a molecular beacon probe to prepare the functional nucleic acid probe; wherein the end of the first DNA single strand and the end of the second DNA single strand are complementary to each other; and the molecular beacon probe comprises an aptamer sequence of foodborne pathogenic bacteria. The fluorescence polarization method based on the functional nucleic acid probe provided by the application has the advantages of high sensitivity, strong specificity, short detection time and the like, and can be used for self-immediate monitoring of the content of foodborne pathogenic bacteria in drinking water or the content of foodborne pathogenic bacteria in environmental water, and has great application value and market value in family health and environmental protection and the like.

This invention discloses a field-effect transistor biosensor based on a framework nucleic acid-gold nanoparticle composite structure and its fabrication method, comprising the following steps: 1) providing a carbon nanotube field-effect transistor substrate; 2) depositing a hafnium oxide dielectric layer on the surface of the carbon nanotube channel; 3) depositing gold nanoparticles on the surface of the hafnium oxide dielectric layer; 4) preparing a tetrahedral framework nucleic acid probe; 5) adding the probe solution to the surface of the gold nanoparticles for incubation and fixation; 6) sealing unreacted sites on the surface of the gold nanoparticles and washing with buffer solution. This invention achieves precise interface modification through atomic layer deposition, electron beam evaporation, and gold-sulfur bond anchoring. It utilizes the rigid structure of the tetrahedral framework nucleic acid to achieve precise, directional, and spaced probe distribution, effectively overcoming Debye shielding limitations and reducing non-specific adsorption. The sensor exhibits ultra-high sensitivity, excellent stability and reusability, strong specificity, and good versatility, and can be used for highly sensitive and specific detection of various targets such as nucleic acids and proteins.

The invention discloses a micro-nano robot based on a DNA tetrahedron nucleic acid probe and magnetic driving and a preparation method and application of the micro-nano robot. A DNA tetrahedron probe (DTP) constructed by a DNA tetrahedron framework (DTH), a double-hairpin structure (H1 and H2) and a double-strand recognition module (dsDNA) is adopted, signal amplification detection can be performed on two breast cancer subtype related miRNAs, namely miR-21 and miR-31, and the probe is combined with magnetic nanoparticles, so that active mixing and enrichment of the probe and a target are realized under the action of an external rotating magnetic field, and the detection sensitivity is high. And through dual-channel fluorescence logic output, the kit has the advantages of high sensitivity, high specificity, short reaction time, visual typing, good biocompatibility and the like.

The application belongs to the technical field of RNA rapid detection, and particularly relates to a rapid amplification and lateral chromatographic detection method for virus RNA and application. The crude virus nucleic acid extract is subjected to a single-tube composite enzyme reaction system reverse transcription, restriction endonuclease digestion and one-way isothermal strand displacement amplification, and then a large amount of specific sequence single-stranded DNA with a marker is generated; in the lateral chromatographic process, the single-stranded DNA is captured by a nucleic acid probe fixed on a detection area of a virus RNA detection test paper, at the same time, avidin labeled on a colloidal gold particle is combined with biotin labeled on the fixed single-stranded DNA, and the colloidal gold particle is indirectly fixed to show a color indication result. The method provided by the application is simple to operate, does not need complicated nucleic acid extraction, does not need large equipment in the whole detection process, and the result can be judged by naked eyes, so that on-site rapid detection can be realized, and the method has the advantages of simplicity, rapidness, high sensitivity and low cost.

The invention relates to a biomarker combination comprising an NF-kappa B activation marker and a nucleic acid probe or primer for determining the mRNA expression levels of one or more genes of BCL2A1, CCL3, G0S2, IL1beta and TNF alpha, the expression levels of at least three of said genes being significantly increased relative to a reference control; the invention also relates to a detection kit, which comprises a reagent for detecting the NF-kappa B activation marker and a nucleic acid probe or primer for determining the gene combination expression level of the BCL2A1, CCL3, G0S2, IL1beta and TNF alpha mRNA. In addition, the invention relates to a method for preparing the detection kit by using a computer program. According to the invention, the activation state of an NF-kappa B signal channel is combined with the expression characteristics of specific inflammation-related genes for the first time, and a stable and consistent ASD molecular diagnosis system is constructed.

The application belongs to the technical field of biological detection, and discloses a multiplex signal amplification system for direct detection of DNA / RNA mutation and application thereof, and particularly discloses a multiplex signal amplification system, which comprises an enzyme-labeled plate detection platform, a first signal amplification platform, a second signal amplification platform and a third signal amplification platform; the enzyme-labeled plate detection platform contains a capture nucleic acid probe; the first signal amplification platform contains a nucleic acid probe complementary to a part of the mutation DNA / RNA to be detected; the second signal amplification platform contains two single-stranded DNA probes; the third signal amplification platform contains streptavidin-labeled horseradish peroxidase; and the nucleic acid probe complementary to the part of the mutation DNA / RNA to be detected is a nucleic acid probe containing a hairpin structure. The multiplex signal amplification system can realize detection of a target mutation gene and has good sensitivity and specificity.