55 results about "Locked nucleic acid" patented technology

The invention relates to the technical field of biology, in particular to a cis-trans typing method for drug-resistant gene mutation EGFR T790M / C797S. The invention provides an EGFR (Epidermal Growth Factor Receptor) T790M / C797S cis-trans typing method based on a locked nucleic acid LNA (Low Nucleic Acid) probe, ultrahigh-specificity DNA (Deoxyribose Nucleic Acid) polymerase and a primer probe combination. The typing method can be used for accurately distinguishing EGFR T790M / C797S double-mutation cis-configuration and trans-configuration; meanwhile, the kit has the advantages of high sensitivity, high specificity, simple steps and low cost, is suitable for detecting low-abundance samples such as circulating tumor DNA (ctDNA) and can meet the clinical rapid detection requirement.

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 preparation method of the locked nucleic acid intermediate D-ribofuranose derivative, provided by the invention, is simple in steps, stable in process, high in selectivity and easy for industrial production, and has a relatively good application prospect. According to the preparation method, the yield and the purity can be greatly improved, the economic cost is reduced, steps are shortened, multiple operation modes are simplified, the amount of organic waste liquid is remarkably reduced, materials and process conditions with safety risks are eliminated, operation with serious equipment loss is avoided, and the process conditions and the result are stable.

The invention provides a primer probe, a kit and a method for identifying A1 / A2-beta casein in milk and dairy products. The primer probe comprises an allele specific primer A1A2-F, an allele specific primer A1A2-R, a probe A2-P and a probe A1-P. The invention further discloses a kit for detecting the allele specific primer A1A2-F, the allele specific primer A1A2-R and the probe A1-P. According to the present invention, based on the 67th single nucleotide polymorphism (SNP) of the ruminant A1 / A2-beta casein gene (CSN2), the gene typing method with characteristics of high specificity and high sensitivity is constructed; according to the method, an ARMS-PCR technology, a locked nucleic acid (LNA) modification strategy and an MGB probe signal detection system are integrated, so that the A1 / A2 allele is accurately distinguished.

This disclosure relates to the modified 3' region elements that confer improved therapeutic properties including, but not limited to, increased stability, increased expression, and / or a reduced innate immune response when introduced into a population of cells, even in the absence of a 5'-cap. The disclosure also provides polynucleotides (e.g., RNA molecules) containing modified 3' regions. In some embodiments of the disclosure, the modified 3' regions contain modified adenosines (e.g., 2'-0-methoxy- adenosine, 2'-fluoro-adenosine, or locked nucleic acid (LNA)-adenosine) and / or a modified thymidine (e.g., inverted deoxythymidine). In some embodiments of the disclosure, the modified 3' region may contain one or more modified internucleoside linkages (e.g., a phosphorothioate linkage) or modified nucleobase (e.g., N1-methyl-pseudouridine). The polynucleotides of the disclosure may be used to express a desired polypeptide in a subject, such as a subject having a disease or condition associated with a deficiency in the corresponding endogenous polypeptide.

The present application relates to the field of biotechnology and molecular detection, in particular to a plant and pathogenic bacteria interaction gene expression analysis method based on UMI tag, comprising: obtaining interaction sample total nucleic acid extract; and constructing a molecular hybridization system with host-specific chimeric blocking probes containing locked nucleic acid modification and 3' end dideoxy modification, and reverse transcription primers; through temperature rising and denaturation and temperature reduction and annealing, the blocking probes are combined with host high abundance nucleic acid to form a hybridization blocking complex, and the primers are combined with pathogenic bacteria nucleic acid; template switching oligonucleotides with unique molecular identifier sequences and reverse transcriptase are added, the host extension is blocked, and UMI is introduced into the pathogenic bacteria complementary chain, and an interaction expression library is generated; after sequencing, the pathogenic bacteria effective reads and the host invalid reads are counted, and the effective interaction signal signal-to-noise ratio is output; the present application can preferentially capture pathogenic bacteria transcripts in the high background of host RNA.

Disclosed herein include systems, methods, compositions, and kits for prevention of antibody oligonucleotide extension. There are provided, in some embodiments, blocking oligonucleotides. In some embodiments, the blocking oligonucleotide comprises a binding region capable of hybridizing barcoding oligonucleotides to form a protected duplex. The blocking oligonucleotide can be a locked nucleic acid (LNA), a peptide nucleic acid (PNA), a DNA, an LNA / PNA chimera, an LNA / DNA chimera, or a PNA / DNA chimera. In some embodiments, the 3′ end of a blocking oligonucleotide is incapable of being extended by a reverse transcriptase or a polymerase. In some embodiments, the blocking oligonucleotide of a protected duplex is incapable of being removed by the strand displacement activity of a reverse transcriptase or a polymerase.

The invention discloses a mutant gene enrichment method, a mutant gene detection method and a mutant gene detection kit based on Cas9 specific cleavage and isonucleotide magnetic beads. The method comprises the following steps: firstly, extracting free DNA; then identifying and cutting a completely matched wild type sequence by using Cas9 protein and specific sgRNA, and reserving the mutant DNA due to base mismatch; cas9 treated DNA and streptavidin magnetic beads containing a biotinylated oligonucleotide probe are incubated, guanine and cytosine in the probe are substituted by isoguanine and isocytosine respectively, and adenine and thymine are modified by locked nucleic acid, so that high-stability homodromous pairing is realized, and mutation DNA is selectively enriched; and finally, carrying out qPCR or sequencing detection on the enriched DNA. The kit has the characteristics of simplicity and convenience in operation, high specificity and high sensitivity, can be used for detecting single-base-level mutation, and is suitable for liquid biopsy, tumor early screening and genetic disease mutation detection.

To provide a C6'-substituted locked nucleic acid-modified cap analog and a method of use thereof.SOLUTION: The present disclosure provides a compound represented by a formula (I), or a stereoisomer, a pharmaceutically acceptable salt or a solvate thereof. In the formula, X is O, S, or the like; R1 to R6 are each H, substituted / unsubstituted C1-C6 alkyl, C2-C6 alkenyl, or the like; Y1a to Y1f are each O, S, CH2, CCl2, or the like; Y2a to Y2d are each =O or =S; Y3a to Y3d are each OH or SH; and B1 and B2 are each a natural / modified pyrimidine nucleotide base or natural / modified purine nucleotide base.SELECTED DRAWING: Figure 4

This invention relates to the field of microbial detection technology, specifically to a precise LNA-qPCR detection method for *Bifidobacterium longum* subsp. *infantii* CICC 6069, as well as specific primers, probes, kits, and their applications for this method. This invention proposes the application of the nucleotide sequence shown in SEQ ID NO:1 in the precise identification and quantitative detection of *Bifidobacterium longum* subsp. *infantii* CICC 6069. By constructing a pan-genome of *Bifidobacterium longum*, mining SNP sites unique to CICC 6069, and designing specific probes SEQ ID NO:14 and primers SEQ ID NO:4 and SEQ ID NO:5 containing locked nucleic acid (LNA) modification, combined with real-time quantitative PCR technology, rapid and precise quantitative detection of this strain at the in vitro level is achieved. This method has advantages such as high specificity, high sensitivity (limit of quantification approximately 500 CFU / mL), and short detection cycle, and is suitable for quality control and quantitative analysis of samples containing CICC 6069, including food, daily chemical, and compound microbial agents.

The application discloses a method for detecting thyroid cancer driving gene mutations based on multiplex fluorescence PCR, and belongs to the technical field of molecular biology detection. The method simultaneously detects nine mutation sites, namely BRAF V600E, NRAS Q61R, NRAS Q61K, HRAS Q61R, HRAS Q61K, KRAS G12V, KRAS G12D, TERT C228T and TERT C250T in a single tube reaction. The method enhances the distinguishing ability of wild type and mutant types by using a locked nucleic acid modified allele-specific primer, suppresses wild type amplification by using a peptide nucleic acid clamp, introduces a competitive internal reference probe to correct amplification efficiency difference, and reduces the sample usage to one fifth of that of a traditional multi-tube scheme. The mutation detection lower limit reaches 0.5% allele frequency, and the total time consumption is not more than 2.5 h.

The present invention discloses a nucleic acid probe combination for detecting rifampicin resistance gene mutation of Mycobacterium tuberculosis, comprising: a plurality of probes for a nucleic acid amplification reaction, each of the plurality of probes being completely complementary to a corresponding one of a plurality of mutant sequences in a drug resistance determining region of an rpoB gene, and at least one of the plurality of probes has a locked nucleic acid (LNA) modification.

The invention discloses a double-end modified probe for detecting single nucleotide polymorphism as well as a design method and application of the double-end modified probe, and through the synergistic effect of thio-modification and locked nucleic acid modification and in combination with SNP sites and 3 '-end artificial mutation design, the distinction degree and detection sensitivity of the probe to wild types and mutant types are improved. Experiments show that the probe can detect a mutation proportion as low as 0.1%, the melting temperature difference between a wild type and a mutant type is greater than 4 DEG C, and the probe is suitable for multiple detection of drug-resistant genes or strain identification of pathogenic microorganisms such as mycobacterium tuberculosis and the like.

An oligonucleotide probe for detecting a genomic alteration includes a first complementary region, a second complementary region, and a semi-complementary region. The first complementary region includes at least three nucleotides that are completely complementary to a mutant allele of a portion of a genomic sequence, the includes at least three nucleotides that are completely complementary to the mutant allele and to the wild type allele, and the semi-complementary region is adjacently flanked by the first complementary region and the second complementary region. The nucleotides of the semi-complementary region are completely complementary to the mutant allele and are not completely complementary to the wild type allele, the first complementary region comprises a first locked nucleic acid adjacent to the semi-complementary region, the second complementary region comprises a second locked nucleic acid adjacent to the semi-complementary region, and the semi-complementary region comprises a third locked nucleic acid.

The present invention relates to RNA molecules, and in particular, but not exclusively, to unlocking nucleotides (UNAs) and unlocking nucleic acid (UNA)-RNA molecules themselves, as well as to methods of making UNA-RNA molecules, and to methods of translating UNA-RNA molecules into proteins. The present invention relates to an improved method of forming UNA-RNA by in vitro transcription, and to the resulting UNA-RNA molecules. Furthermore, the present invention relates to a novel method for enhancing the expression and / or translation of RNA, i.e. Protein expression, as well as a method for improving the stability of RNA molecules. The invention also relates to reducing the activation of innate perception, interferon production, and / or degradation of RNA molecules in a host. The invention also relates to the use of said UNA-RNA molecules in vaccines and other therapeutic pharmaceutical compositions, as well as their use in immunization and therapy, such as RNAi, gene therapy, gene editing and protein substitution. Furthermore, the present invention relates to a novel method of enhancing RNA translation, i.e. Protein expression.

The invention belongs to the technical field of biological detection, and particularly relates to a DNA polymerase activity detection method, a detection kit and application thereof. The method is based on a qPCR technology, and efficient and accurate detection of DNA polymerase activity is realized by designing a special reaction system and optimizing an experimental process. In the incubation stage, an oligonucleotide fragment which is annealed in advance and contains mismatched basic groups is used as a primer and a template of DNA polymerase to be detected to carry out polymerization reaction; in a detection stage, a polymerization reaction product is quantified by virtue of a qPCR technology through a locked nucleic acid enhanced TaqMan probe competitive binding strategy. The method is simple and convenient to operate, good in safety, high in sensitivity and wide in linear range, and can be applied to the scenes of activity detection of different types of DNA polymerase, enzyme preparation screening, hot start polymerase performance evaluation, polymerase activity quantification in complex samples and the like, so that the method has good practical application value.

The present application relates to the technical field of biological detection, and a double-path apple codling moth identification system integrating RPA-LFS rapid screening and TaqMan qPCR real-time quantification, a locked nucleic acid probe primer set, a kit and a method thereof. COI Through gene screening and primer screening, the present application designs recombinase-mediated isothermal amplification (RPA) and TaqMan qPCR primers and probes for the barcode of the cytochrome oxidase subunit I (COI) gene of apple codling moth, and establishes an RPA-LFS method in combination with immunolateral flow. The RPA-LFS method can complete amplification and visual judgment within 20 minutes at 39 DEG C, and is suitable for on-site rapid detection. The TaqMan qPCR method can realize high-sensitivity quantitative analysis under the condition of crude extract. Through double-path complementary design, the consistency of the results of on-site screening and laboratory confirmation detection is realized, and the method is simple to operate and suitable for pest quarantine at ports, epidemic monitoring in agricultural production areas and quarantine at storage and transportation links.

The invention discloses a composition for detecting type I and type II monkey pox virus typing and a typing detection kit and application thereof, the composition comprises an LNA-Taqman probe for detecting the monkey pox virus I, the nucleotide sequence of the LNA-Taqman probe is shown as SEQ ID NO. 1, the nucleotide sequence of the LNA-Taqman probe is shown as SEQ ID NO. 2, and the nucleotide sequence of the LNA-Taqman probe is shown as SEQ ID NO. The nucleotide sequence of the upstream primer for typing detection of the monkey pox virus is shown as SEQ ID NO. 2; and a monkey pox virus typing detection downstream primer, the nucleotide sequence of which is as shown in SEQ ID NO. 3. According to the invention, the characteristic that the hybridization stability is sharply reduced when a locked nucleic acid (LNA) probe is mismatched with a single base is utilized, and an LNA monomer is accurately designed at a monkey pox virus typing SNP key site of the probe. By means of the design, typing signal differences can be remarkably amplified, and highly homologous virus branches can be effectively distinguished. The problems of cross reaction and false positive caused by high mismatching tolerance of a traditional DNA probe are avoided, and the accuracy of typing identification is ensured.

The invention relates to the technical field of biological medicines and functional nucleic acid molecules, and discloses pearl mussel source miRNA and application thereof, a biological preparation and application thereof. The pearl mussel source miRNA is miRNA derived from a pearl mussel pallium exosome, contains a sequence shown in SEQ ID NO: 1, and can be modified by phosphorylation, locked nucleic acid and the like to enhance the stability. The biological preparation containing the miRNA can be used for preparing a medicine for preventing and / or treating osteoporosis or functional food for promoting bone mineralization, and the bone mineral density and the bone microstructure are improved by promoting osteoblast proliferation, differentiation and mineralization. According to the invention, marine biological resources which are natural in source and high in safety are utilized, and new active ingredients and technical schemes are provided for prevention and treatment of osteoporosis.

A computer-implemented framework for designing high-efficiency and high-specificity nucleic acid molecules targeting transfer RNA (tRNA) and its derivatives (tDRs). The framework comprises two primary algorithms. The first, tBOND-G, designs guide RNAs (gRNAs) for Cas13-mediated tRNA cleavage by calculating physical parameters, including target site accessibility and binding energy, and processing them through a Support Vector Machine (SVM) model to predict cleavage efficiency. The second algorithm, tBOND-L, designs therapeutic Locked Nucleic Acid-modified antisense oligonucleotides (LNA-ASOs) that specifically target a tDR without binding to its parent tRNA. This method utilizes a processor to derive an efficiency score based on relative binding affinity and a specificity score based on simulated competitive binding environments.

The present invention relates to RNA molecules and, in particular, but not exclusively, to methods of making RNA molecules, and methods of translating RNA molecules into proteins. The present invention relates to improved methods of forming UNA-RNA by in vitro transcription, and the resulting RNA molecules. Furthermore, the present invention relates to novel methods of enhancing the expression and / or translation of RNA, i.e. protein expression, and methods for improving the stability of RNA molecules. The present invention also relates to reducing the activation of innate sensing, interferon production, and / or degradation of RNA molecules in a host. The present invention also relates to the use of said RNA molecules in vaccines and other therapeutic pharmaceutical compositions, and their use in immunization and therapy, such as RNAi, gene therapy, gene editing and protein replacement. In particular, the present invention relates to locked nucleotide and locked nucleic acid (LNA)-RNA molecules, and the use or incorporation of such locked nucleotide and LNA-RNA molecules in any of the above mentioned applications.

The invention provides a probe and a kit for detecting EB (Epstein-Barr) virus and application of the probe and the kit. The probes comprise a probe specifically combined with an EBER1 gene and a probe specifically combined with an EBER2 gene; the nucleotide sequence of the probe is shown as any one of SEQ ID NO: 1 to SEQ ID NO: 18. On the basis of the probe, multi-Dig labeling and locked nucleic acid modification are carried out, so that the developing intensity, the probe hybridization efficiency and the stability are improved. On the basis, the probes are combined for use, after combined use, the developing background is further reduced, the specificity is enhanced, and the detection effect is further improved.

The invention discloses a digital PCR detection system, kit and method for detecting single nucleotide polymorphism and copy number variation of a CYP2D6 gene and application, and belongs to the technical field of molecular biology. The method comprises the following steps: designing three pairs of CYP2D6 and reference gene RPP30 specific primers, matching with four locked nucleic acid modified TaqMan fluorescent probes, constructing a single-tube multiple digital PCR detection system, and synchronously and accurately detecting three single nucleotide polymorphisms of CYP2D6 * 10, CYP2D6 * 36 and CYP2D6 * 41 and CYP2D6 * 5 copy number variation; wherein the RPP30 reference gene can synchronously realize sample amplification quality control and target gene absolute quantitative calibration. The detection system depends on the advantages of high specificity of the LNA probe and high sensitivity of digital PCR, the detection sensitivity reaches 0.02 ng / mu L, the detection period is short, and the result is stable and reliable.

The invention discloses a monkey pox virus typing nucleic acid detection primer probe combination, a kit and application, and belongs to the technical field of biology. The primer probe combination comprises six groups of specific primer pairs and six corresponding probes, and the six groups of specific primer pairs and the six corresponding probes are respectively used for accurately identifying the monkey pox viruses such as Clade Ia, Clade Ib, Clade II, Clade IIb A.1, Clade IIb B.1 and Clade IIb C.1; the primer probe combination provided by the invention has the advantages of high specificity, high sensitivity and accurate typing, and effectively overcomes the influence of target sequence deletion or mutation caused by virus variation on the detection accuracy through combination of locked nucleic acid modified probes and a multiplex fluorescent PCR technology; the method can realize rapid and parallel identification of main branches and epidemic pedigree of the monkey pox virus, and provides reliable technical support for epidemic prevention and control and clinical diagnosis.

The present invention generally relates to tRNA-based effector molecules containing a lock nucleic acid (LNA) moiety, as well as compositions and related methods. This disclosure features modified tRNA-based effector molecules (TREMs, e.g., TREM or TREM fragments), as well as related compositions and uses. As provided herein, TREMs are complex molecules capable of mediating various cellular processes. TREMs disclosed herein include, for example, nucleotides containing a lock nucleic acid moiety or a 2',5'-linked nucleotide on a component nucleotide (e.g., a nucleotide sugar).

The present invention discloses a locked nucleic acid derivative (formula II) allyl double bond migration preparation method, the method is characterized in that terminal group olefin represented by a formula I is subjected to a reaction under catalysis of a transition metal complex and an additive to obtain internal olefin represented by the formula II, the structures of the formula I and the formula II are as shown in the specification, and R1, R2 and B are defined in the specification. According to the reaction, the conversion rate of the raw materials is increased, the cost input is reduced, the cyano-locked nucleic acid is more convenient to produce in a quantitative manner, and the method has a better application prospect.

The present disclosure includes systems, methods, compositions, and kits for preventing antibody oligonucleotide extension. In some embodiments, a blocking oligonucleotide is provided. In some embodiments, the blocking oligonucleotide comprises a binding region capable of hybridizing to a barcoded oligonucleotide to form a protected duplex. The blocking oligonucleotide can be a locked nucleic acid (LNA), a peptide nucleic acid (PNA), a DNA, an LNA / PNA chimera, an LNA / DNA chimera, or a PNA / DNA chimera. In some embodiments, the 3' end of the blocking oligonucleotide is not extendible by a reverse transcriptase or a polymerase. In some embodiments, the blocking oligonucleotide of the protected duplex is not removable by the strand displacement activity of a reverse transcriptase or a polymerase.

The application discloses a primer pair, a kit and application for detecting a BCR-ABL1 gene T315I mutation. The third and / or fourth bases of the 3' end of the upstream primer of the primer pair are introduced with corresponding mismatched bases, the second base of the 3' end of the primer is introduced with a dealkylation site, and the terminal base of the 3' end of the primer is modified by a locked nucleic acid. The primer pair and the kit for detecting the BCR-ABL1 gene T315I mutation can selectively amplify the T315I mutation, realize high-sensitivity detection (up to 10 copies) of the T315I mutation, have good specificity and high precision, can effectively distinguish the wild type and the mutant site, can eliminate the interference of the adjacent site T315A, are simple and rapid to operate, have a short time consumption in the whole detection process, can obtain a detection result within 90 minutes, are high in detection efficiency and low in cost.

The invention relates to the technical field of biology and the technical field of food safety detection, and particularly discloses a nucleic acid aptamer aiming at insecticide imidacloprid, a derivative of the nucleic acid aptamer and application of the nucleic acid aptamer. The nucleic acid aptamer has a DNA sequence as shown in SEQ ID NO: 1, and covers variants of which the homology is greater than or equal to 90%, chemically modified derivatives, and locked nucleic acid, peptide nucleic acid or thiophosphate nucleic acid formed by skeleton modification. The aptamer has high specificity and high affinity to imidacloprid, is suitable for the fields of agricultural product residue detection, environmental monitoring, biosensor construction and the like, and is especially suitable for rapid and on-site screening of imidacloprid. The aptamer has the advantages of being simple and convenient to synthesize, low in cost, good in stability, easy to modify and the like, can be used for preparing a detection probe, and is adaptive to various sensing platforms such as color comparison, fluorescence, electrochemistry and the like.

The invention discloses SART1 gene antisense oligonucleotide and application thereof in tumor treatment, and relates to the technical field of biological medicine. The sequence of the antisense oligonucleotide of the SART1 gene provided by the invention is as shown in any one of SEQ ID NO.1-3, or the sequence of the antisense oligonucleotide of the SART1 gene has at least 90% sequence identity with the sequence as shown in any one of SEQ ID NO.1-3; three nucleosides from the 5'end or three nucleosides from the 3 'end of the sequence of the SART1 gene antisense oligonucleotide are all modified by locked nucleic acid.The SART1 gene antisense oligonucleotide salt provided by the invention can reduce mRNA transcription of the SART1 gene and / or promote mRNA degradation of the SART1 gene, so that the SART1 gene antisense oligonucleotide can be used for anti-tumor treatment.