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62 results about "Single-stranded binding protein" patented technology

Single-stranded binding proteins (SSBs) are a class of proteins that have been identified in both viruses and organisms from bacteria to humans.

In vitro recombination method

The present invention relates, e.g., to in vitro method, using isolated protein reagents, for joining two double stranded (ds) DNA molecules of interest, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule share a region of sequence identity, comprising contacting the two DNA molecules in a reaction mixture with (a) a non-processive 5′ exonculease; (b) a single stranded DNA binding protein (SSB) which accelerates nucleic acid annealing; (c) a non strand-displacing DNA polymerase; and (d) a ligase, under conditions effective to join the two DNA molecules to form an intact double stranded DNA molecule, in which a single copy of the region of sequence identity is retained. The method allows the joining of a number of DNA fragments, in a predetermined order and orientation, without the use of restriction enzymes.
Owner:TELESIS BIO INC

Western blot kit for detecting antibody of autoimmune disease and preparation method thereof

ActiveCN103105489AOvercome the cumbersome operation of individual detection one by oneImprove accuracyMaterial analysisAntigenAnti-mitochondrial antibody
The invention provides a western blot kit for detecting the antibody of autoimmune disease and a preparation method of the western blot kit, and relates to a western blot kit for detecting related antibodies of various autoimmune diseases, aiming at overcoming the technical defect that a western blot product is unavailable for testing and screening various autoimmune diseases in the prior art. The nitrocellulose membrane or the nylon membrane contains at least two parallel detection lines coated by at least two of ten natural antigens or recombinant antigens, i.e. dsDNA (deoxyribonucleic acid), Sm / RNP (ribonucleoprotein), CCP (critical compression pressure), SSA (sulfosalicylic acid), SSB (single-strand binding protein), GAD (glutamic acid decarboxylase), ICA (islet cell antibody), IA-2A (islet cell), TG (triglyceride) and AMA-M2 (anti-mitochondrial antibody), a high-concentration quality control band, a median-concentration quality control band and a low-concentration quality control band. The deficiency of the detection sensitivity and the specificity of the single autoantibody can be overcome, the operating complexity for independently detecting the related autoantibody of various diseases one by one can be overcome, and the detection efficiency and the result judging accuracy degree can be greatly improved.
Owner:SHENZHEN YHLO BIOTECH

Normal-temperature and constant-temperature nucleic acid amplification method using fluorescence probe

The invention relates to a normal-temperature and constant-temperature nucleic acid amplification method using a fluorescence probe. The normal-temperature and constant-temperature nucleic acid amplification method includes: under constant temperature, single-strand binding protein partially opens the double-strand parental strand into a single strand; recombinant enzyme is combined with primer to form complex, the complex is combined to the parental strand under the action of auxiliary protein, and the fluorescence probe is combined with a complementation area; DNA polymerase is combined to the 3' tail end of the primer to perform sub-strand extension; exonuclease identifies the tetrahydrofuran locus on the fluorescence probe under a double-strand state, fluorescent groups and quenching groups are separated after enzyme digestion to release fluorescence; after the fluorescence probe is cut off, the 3'-OH end originally sealed by probe 3' tail end modification is exposed, and DNA polymerase can continuously extend to form a sub-strand; qualitative and semi-quantitative detection can be performed on a to-be-detected sample by detecting the shape of an amplification curve and the strength of a fluorescence signal. The nucleic acid amplification method has the advantages that normal-temperature and constant-temperature detection can be performed, and qualitative and semi-quantitative detection can be performed on nucleic acid to-be-detected objects.
Owner:ZHEJIANG SHANCE HEQISHI BIO SCI& TECH CO LTD

Helicase-dependent isothermal DNA amplification-based method for detecting microRNA

The invention discloses a helicase-dependent isothermal DNA amplification-based method for detecting microRNA. The method comprises the following steps: (1) extracting total RNA in a sample; (2) adding a single-strand DNA probe and excessive exonuclease I into the extracted total RNA, incubating in a reacting buffer solution to realize specific combination of target microRNA and the single-strand DNA probe, and removing residual single-strand DNA probe by utilizing the excessive exonuclease I; and (3) adding upstream primer, downstream primer, single-strand binding protein and helicase to the reaction system, amplifying the target microRNA, and detecting expression of the target microRNA through a fluorescent signal. According to the method, the background is reduced by digesting the exonuclease I, and the signal is amplified by means of exonuclease assisted isothermal amplification (HDA) reaction, so that the microRNA can be quickly and sensitively detected.
Owner:SHANDONG NORMAL UNIV

RPA method for detecting 3 type human adenovirus, special primer, probe and application thereof

The invention provides a RPA detection method for detecting 3 type human adenovirus, a special primer, a probe and an application thereof in detecting 3 type human adenovirus. The detection method, the special primer and the probe thereof are designed on the basis of 3 type human adenovirus hexon gene conserved sequence, with oligonucleotide sequences shown as SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4. According to the invention, a novel isothermal amplification technology RPA is firstly applied to the detection of 3 type human adenovirus. According to the method, an enzyme reaction process ofin vivo DNA replication is simulated; amplification of DNA template relies on a specific combination of enzyme and protein (recombinase, single-stranded binding protein and DNA polymerase); amplification of specific nucleotide sequence is realized under a constant temperature at 37 DEG C; amplified products can be visually identified through a lateral chromatography test strip. The detection method established by the invention has the advantages of high sensitivity, limit of detection reaching up to 101 copies / reaction, high specificity, no cross reaction with other pathogens, high detection speed, only 20min reaction time, simple and convenient operation, and suitability for onsite detection.
Owner:李越希

RPA method for detecting 21 type human adenovirus, special primer, probe and application thereof

The invention provides a RPA detection method for detecting 21 type human adenovirus, a special primer, a probe and an application thereof in detecting 21 type human adenovirus. The detection method,the special primer and the probe thereof are designed on the basis of 21 type human adenovirus hexon gene conserved sequence, with oligonucleotide sequences shown as SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4. According to the invention, a novel isothermal amplification technology RPA is firstly applied to the detection of 21 type human adenovirus. According to the method, an enzyme reaction processof in vivo DNA replication is simulated; amplification of DNA template relies on a specific combination of enzyme and protein (recombinase, single-stranded binding protein and DNA polymerase); amplification of specific nucleotide sequence is realized under a constant temperature at 37 DEG C; amplified products can be visually identified through a lateral chromatography test strip. The detection method established by the invention has the advantages of high sensitivity, limit of detection reaching up to 102 copies/reaction, high specificity, no cross reaction with other pathogens, high detection speed, only 20min reaction time, simple and convenient operation, and suitability for onsite detection.
Owner:李越希

RPA (recombinase polymerase amplification) method, PRA special primer and RPA probe for detecting universal human adenovirus, and application of RPA probe

The invention provides an RPA (recombinase polymerase amplification) method, an PRA special primer and an RPA probe for detecting universal human adenovirus, and application of the PRA special primerand RPA probe in detection of the universal human adenovirus. The detection method, the special primer and the probe are designed based on a common conserved sequence of type 3, type 7, type 11, type21 and type 55 human adenovirus hexon genes, and have the oligonucleotide sequences shown as SEQ ID NO. 3, SEQ ID NO. 4 and SEQ ID NO. 5. According to the method, the novel isothermal amplification technology is applied to detection of the universal human adenovirus for the first time; the enzymatic reaction process of in-vivo DNA replication is simulated, a specific enzyme and protein combination(recombinase, single-stranded binding protein and DNA polymerase) is used for DNA template amplification, amplification of a specific nucleotide sequence can be realized at a constant temperature of37 DEG C, and amplification products can be visually judged through a lateral chromatographic test strip. The detection method has high sensitivity, and the detection limit can reach 101 copies / reaction; the specificity is high, and no cross-reaction with other pathogens exists; the detection speed is high, and the method is simple and convenient to operate.
Owner:李越希

Constant-temperature detection method and kit for instantly detecting listeria monocytogenes

The invention discloses a constant-temperature detection method and kit for instantly detecting listeria monocytogenes. The constant-temperature detection kit does not depend on high-energy molecules such as ATP and phosphocreatine. Compared with the existing constant-temperature amplification system, the constant-temperature detection kit is relatively simple in components, relatively low in cost and relatively convenient to use. Key components comprise dNTPs, polymerase Bsu, single-strand binding protein and recombinase (E.coli Rect). The constant-temperature detection kit can be matched with a fluorescently-labeled probe based on a constant-temperature amplification reaction reagent to carry out instant fluorescence detection. The constant-temperature amplification system is applied to amplification of complex templates; amplification is rapid and stable; the amplification reaction can be completed in 30 minutes; amplification is high in sensitivity and accuracy; the possibility of mismatching is low; the amplification effect is excellent.
Owner:GUANGZHOU HEAS BIOTECH CO LTD

Preparation method, expression gene, recombinant expression vector and application of gp32 single-strand binding protein

The invention relates to a preparation method, expression gene, recombinant expression vector and application of gp32 single-strand binding protein. The expression gene of the gp32 single-strand binding protein comprises a nucleotide sequence shown in SEQ ID No.1. The expression gene of the gp32 single-strand binding protein can achieve a large amount of soluble expression in escherichia coli, andthe purity of the expressed gp32 single-strand binding protein is greater than 95%.
Owner:深圳市艾伟迪生物科技有限公司
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