325 results about "Proteinase K" patented technology

A highly conserved, immunologically accessible antigen at the surface of Neisseria meningitidis organisms. Immunotherapeutic, prophylactic and diagnostic compositions and methods useful in the treatment, prevention an diagnosis of Neisseria meningitidis diseases. A proteinase K resistant Neisseria meningitidis surface protein having an apparent molecular weight of 22 kDa, the corresponding nucleotide and derived amino acid sequences (SEQ ID NO: 1, NO:3, NO:5 and NO:7: SEQ ID NO: 2, NO:4, NO:6, and NO:8), recombinant DNA methods for the production of the Neisseria meningitidis 22 kDA surface protein, and antibodies that bind to the Neisseria meningitidis 22 kDA surface protein.

The invention belongs to the field of molecular biology and particularly relates to a kit for extracting ribonucleic acid (RNA) by virtue of a paramagnetic particle method and an extraction method. The kit contains tissue digestion fluid, lysate, proteinase K, DNase I, DNase IBuffer, nucleic acid, extraction magnetic beads, washing liquid I, washing liquid II and eluant. The invention further discloses a method for extracting the tissue RNA by virtue of the kit. According to the kit and the method, the extraction yield and purity of RNA are increased, the integrity of RNA is improved, meanwhile, the automatic extraction is realized, and the simultaneous parallel testing of multiple samples is realized, so that the labor cost and the time cost are saved.

The invention provides a preservative composition of blood DNA. The preservative composition of blood DNA comprises, by weight, 20 to 50 parts of a preservative, 5 to 15 parts of an anticoagulant, 0.5 to 5 parts of a nuclease inhibitor, 1 to 5 parts of a metabolic inhibitor and 40 to 70 parts of water, wherein the nuclease inhibitor comprises one or more of diatomite, bentonite, ethyl alcohol, formamide, vanadyl-ribonucleoside complex, dithioerythritol, diethyl pyrocarbonate, proteinase-K, heparin, beta-mercaptoethanol, hydroxylamine-O-copper ions, ammonium sulfate, dithiothreitol, cysteine, and 4-formylbenzoic acid and oxovanadium Schiff base complex.

The present invention provides antibiofilm composition comprising two or more agents selected from the group consisting of DispersinB™, 5-Fluorouracil, Deoxyribonuclease I and Proteinase K for preventing growth and proliferation of biofilm-embedded microorganisms in wound care, oral care, and disease-related infections and methods of treatment in mammals. The invention further provides methods for preparing medical devices, and wound care devices using an antibiofilm composition comprising two or more antimicrobial agents selected from the group consisting of DispersinB™, 5-Fluorouracil, Deoxyribonuclease I and Proteinase K.

A new method of simultaneous and separate extraction of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) from a biological sample, indifferently from fresh, frozen, fixed or autoptic tissue with a weight no less than 5 mg. The main steps of the method are: 1/sample lysis in a solution composed of a caotropic agent (urea or guanidine salt), a ionic detergent (SDS or SLS), a proteolytic enzyme (proteinase K, trypsin, chymotrypsin, pepsin or pronase), a reducing agent (β-mercaptoethanol or dithiothreitol); 2/deproteination; 3/precipitation of RNA from aqueous phase; 4/precipitation of DNA from organic phase. The invention includes an extraction kit for nucleic acids, also of viral origin.

Assays are provided for rapid detection, with high specificity of the pathogenic form of prion protein responsible for neurodegenerative diseases affecting humans and animals, such as transmissible spongiform encephalopathy in bovine, sheep, and cats. Also provided are assays for testing animal feedstock, such as animal feed, for the presence or concentration of pathogenic prion protein. Results are available in from about 0.5 to about 20 minutes and preferably within from about 5 to about 10 minutes. The assays employ proteinase-K to remove normal prion protein from a biological sample, so that the sample may be analyzed by immunochromatography to determine the presence and concentration of pathogenic prion protein. Because the proteinase-K is immobilized on a solid support for in situ removal of interfering components, the present invention obviates the need for subsequent extraction of the desired analyte. All aspects of the present invention are suitable for quantifying the minimal detectable amount of pathogenic prion protein in a biological sample. Moreover, the simplicity of sample preparation makes the present invention suitable for use in the field.

The invention discloses a lysate, nucleic acid extraction kit, and nucleic acid extraction method for nucleic acid extraction. The lysate for the nucleic acid extraction is prepared from 0.5 to 2M ofsodium salt, 2 to 5M of guanidine salt, 1 to 5mM of a metal ion complexing agent, 0.25% to 1% of a nonionic surfactant, and 1% to 3% of PEG, and the molecular weight of the PEG is 6000 to 10000 Da, preferably 8000 Da. The lysate has a reasonable ratio, cells can be fully and effectively lysed, the lysis efficiency is high, so that the nucleic acids in the cells can be fully released, and the nucleic acids with higher concentration and purity can be obtained, and the quality and extraction efficiency of the nucleic acids are improved advantageously; and according to the lysate and the kit, digestion and removal of protein can be conducted without the aid of proteinase K during the entire nucleic acid extraction process, alcohols are not usedfor precipitate the nucleic acids,the obtained nucleic acids have high purity, and fragments are complete.

The invention discloses a salt-tolerant ethanol-tolerant protease-tolerant surfactant-tolerant exoinulinase, a gene thereof, a vector and a strain. The exoinulinase InuAJB13 possesses the following properties: the optimum pH is 5.5, 50% or more of enzymatic activity is maintained in the pH scope of 4.0-7.0; the remnant enzyme activity reaches 90% or more after exoinulinase is processed by a buffer with a concentration of 0.1 M and pH of 4.0-7.0 for 1 h; the optimum temperature is 55 DEG C, and exoinulinase has the enzyme activity at 10-70 DEG C; a NaCl solution with a concentration of 0.6-4.5 M is capable of improving the enzyme activity by 0.2-0.6 times; 100% of the enzyme activity can be kept after exoinulinase is processed by a NaCl solution with a concentration of 0.2-4.5 M at 37 DEG C for 60 min; exoinulinase keeps the activity in 10% (V/V) of ethanol; exoinulinase keeps 88% or more of the activity after being processed in 3.0-15.0% (V/V) ethanol at 37 DEG C for 60 min; exoinulinase activity is not influenced or slightly influenced by trypsin, protease K, surfactants, most of metal ions, and commercial liquid laundry detergents; and exoinulinase is capable of hydrolyzing inulin, cane sugar, raffinose, stachyose, beta-2,6-fructan (levan) and soluble starch. The exoinulinase disclosed by the invention is applicable to industries such as feed, foodstuff, washing and biofuels.

The invention discloses a method and a reagent for extraction of viral/bacterial nucleic acid in an animal sample. The method comprises the following steps: taking the animal sample, and adding a lysate for lysis at a room temperature so as to obtain a lysis reaction solution; and adding an ethanol solution into the lysis reaction solution, transferring an obtained mixture into a silica gel containing adsorption membrane centrifugal column, carrying out adsorption with a manner of low-speed centrifugation, and carrying out washing and eluting. According to the invention, the bacterial nucleicacid extracted by using the method provided by the invention has high purity; no proteinase K for sample digestion is needed to be used in the process of extraction; reagent components and an operation flow are simplified; the cost of hardware equipment can be reduced through a manner of low-speed centrifugation in the process of extraction; a low-speed palm centrifuge can be adopted, is convenient to carry and is applicable to on-site operation; the components of the lysate are optimized; the binding ability of nucleic acid molecules to a silica-gel adsorption membrane is reinforced; and goodnucleic acid extraction efficiency is guaranteed.

The invention provides a kit for simultaneously detecting mutations in mitochondria DNA A1555G and C1494T related to maternally inherited drug-induced deafness and a using method thereof. The kit comprises a reagent for extracting sample genomic DNA, a PCR amplification reactive reagent, a primer mixed liquor, a positive control template and a negative control template. The using method of the kit mainly comprises the following steps: using blood, hair with follicle, oral mucosa doctor blade, saliva, and the like, as a sample; adopting a proteinase K digestion pyrolysis method to extract genomic DNA; and then simultaneously detecting mutations in A1555G and C1494T by multiple allele specific PCR. The kit is used for detecting the mutations in mitochondria DNA A1555G and C1494T related to maternally inherited drug-induced deafness and is more rapid, economical and simpler than the single detection for mutation in A1555G or C1494T, and the kit has low requirements for equipment and environment and is conducive to promotion and application.

An aqueous composition comprising a denaturing agent, a chelator, a buffering agent and a protease for the extraction of nucleic acid from a sample of bodily fluid, such as saliva, such that the extracted nucleic acid is stable for at least fourteen days at room temperature and can be directly utilised in an amplification reaction without further processing. In particular, said composition comprises SDS, Cyclohexanediamine tetraacetate, Tris-HCl and proteinase K. A method and kit for the amplification of DNA directly from a bodily fluid, comprising said composition, is further provided.

The invention relates to a deoxyribonucleic acid (DNA) extraction technology, in particular to a method for simply, conveniently and quickly extracting trace total DNA of single roes and fries. The method comprises the following steps of: fixing and preserving collected roes or fries; washing to remove stationary liquid, adding DNA extraction buffer solution and lysis buffer solution, shearing the roes or the fries, adding proteinase K and ribonucleic acid (RNA) enzyme, incubating and digesting; and precipitating protein by using high-concentration NaCl solution, adding isopropanol to precipitate DNA, washing a precipitate by using 70 percent ethanol to remove salt ions, and adding ultrapure water or tris-ethylenediaminetetraacetic acid (TE) solution and dissolving after the ethanol is volatilized. The method solves the problem of the limitation of the application the conventional total DNA extraction method to samples with the low content of genomic DNA, such as roes or fries and thelike.

The invention discloses a method for extracting genomic DNA from giant salamander skin mucus. The method comprises the following steps: 1, saving a sample; 2, replacing ethanol; 3, splitting a cell; 4, precipitating protein; 5 precipiating DNA; 6, washing the DNA; and 7, obtaining DNA, wherein required pyrolysis liquid can be obtained by mixing 160 microliter 1 mol/L nacl aqueous solution, 40 microliter 1 mol/L TRIS-HCI aquenous solution with pH value of 8.0, and 2 microliter 100mmol/L EDTANa2 aqueous solution with pH value of 8.0 uniformly at normal temperature, and then adding 160 microliter 10% of SDS aqueous solution and 20 microliter 10 mg/ml proteinase K aqueous solution to the mixture and quickly mixing the mixture. The method provided by the invention adopts giant salamander skin mucus or skin as an extracting material of the genomic DNA, so that the giant salamander organism will be protected without damage.

The invention discloses a magnetic bead-based DNA extraction method, a lysate and a kit. The lysate is characterized in that a buffer solution with a sodium ion content of greater than or equal to 140mmol and a pH value of 7 to 9 contains a chaotropic agent with a concentration of 3 mol/L to 6 mol/L, 5 mmol to 20 mmol of a metal chelating agent and 5% to 15% by mass of polyethylene glycol. The extraction method comprises the following steps: (1) acquiring a cleavage reaction system; (2) treating the cleavage reaction system; (3) adding nanometer magnetic beads; (4) acquiring magnetic beads binding to DNAs; (5) carrying out washing and then performing DNA elution. The kit comprises the lysate, a proteinase K solution, a magnetic bead dispersion liquid, a first washing solution, a second washing solution, a third washing solution and an eluent. Reagents used in the invention are non-toxic, so experimental safety is improved; and only one-step liquid adding operation is required, so labor cost is greatly reduced.

The invention relates to the application field of nanometer and micrometer materials, in particular to a method for quickly extracting large numbers of genome DNA from biological samples with magnetic particulates. The method comprises the following steps of: (1) cracking biological samples: taking a certain amount of biological samples such as blood, tissue and like, adding proteinase K solution and guanidine hydrochloride cracking liquid with the concentration of 4 to 6 M, evenly mixing, and fully cracking in 56 DEG C water bath for 5 to 20 min; (2) adding combined solution into sample cracking liquid to form mixed solution containing magnetic particulates (DNA compound); (3) separating the magnetic particulates (DNA compound) from the mixed solution; (4) washing; (5) eluting: adding eluting liquid to enable DNA to be eluted from the magnetic particulates, separating the magnetic particulates and the eluted DNA under the action of an exterior magnetic field, and collecting supernatant liquid to obtain rarefied DNA. By using the invention, high purity DNA can be successfully extracted from large numbers of samples, large-scale instruments such as a centrifuge and like are not needed, the operation steps are simple and convenient and need short time.

The invention discloses a quantitative detection kit for human immunodeficiency virus HIV-1. The quantitative detection kit comprises sample treating agent, an upstream primer HIV-F used for target nucleotide amplification, a downstream primer HIV-R used for target nucleotide amplification, a Taqman probe HIV-P for detecting target nucleotide and RNA one-step reaction buffer, wherein the sample treating agent comprises guanidinium isothiocyanate, sodium citrate, dodecyl creatine sodium, beta-mercaptoethanol and proteinase K; a fluorescent group and fluorescence quencher are respectively combined to two ends of the Taqman probe HIV-P. When the quantitative detection kit is used for detecting, a to-be-detected sample is directly mixed with the sample treating agent and then is used for subsequent detection. Compared with a traditional quantitative detection kit for HIV virus, the quantitative detection kit has the advantages that the step of extracting and purifying nucleic acid molecules from the sample is omitted, and time-saving and simple operation is achieved.