61 results about "Nuclease inhibitor" patented technology

Methods and compositions for inhibiting and / or inactivating nucleases by using nuclease inhibitors are provided. The nuclease inhibitors comprise anti-nuclease antibodies and non-antibody nuclease inhibitors.

The present invention concerns a compositions and method for isolating a nucleic acid from a cell-containing sample. There is disclosed a method comprising obtaining at least one cell-containing sample, which comprises a cell containing nucleic acid, obtaining at least one catabolic enzyme, obtaining at least one nuclease inhibitor, preparing an admixture of the sample, the catabolic enzyme, and the nuclease inhibitor, maintaining the admixture under conditions where the catabolic enzyme is active, and agitating the admixture, where the sample is digested to produce a nucleic acid-containing lysate of the sample.

The invention discloses a blood collecting container, and particularly relates to a container for collecting nucleic acid preserved at normal temperature. The container comprises a collecting pipe and a rubber plug, wherein the rubber plug is sealed on a pipe orifice of the collecting pipe; a protective cap is arranged outside the rubber plug; an additive is arranged inside the collecting pipe; and the additive is prepared from a blood anticoagulant and a buffer solution thereof, an antiseptic fixative, a nuclease inhibitor, a metabolic inhibitor and a cell membrane stabilizer in a mixing manner. By adopting the container, the collected sample can be kept in an initial state for 7-14 days or even longer time at normal temperature, and does not decay, so that the detection result is not affected. The collecting container can be applied to collection, storage and transportation of a blood sample in a disaster, a war or a hard area, can be applied to collection, storage and transportation of other tissues, cells and the like in a scientific experiment, and also can be applied to other clinical inspection items. By adopting the container disclosed by the invention, the storage condition after the sample is collected is reduced. Thus, the labor of a worker is reduced, and the work efficiency is improved.

Methods and compositions for inhibiting and / or inactivating nucleases by employing nuclease inhibitors are provided. The nuclease inhibitors comprise anti-nuclease antibodies and non-antibody nuclease inhibitors. The anti-nuclease antibodies of the present invention may be a polyclonal or monoclonal antibodies, and may be anti-ribonuclease antibodies, anti-deoxyribonuclease antibodies, or antibodies to non-specific nucleases. A preferred embodiment comprises at least two nuclease inhibitors, and is referred to as a nuclease inhibitor cocktail. In some specific embodiments, the invention concerns methods of performing in vitro translation comprising obtaining a first nuclease inhibitor, which inhibitor is further defined as an anti-nuclease antibody, and placing the anti-nuclease antibody in an in vitro translation reaction. In many cases, the in vitro translation reaction comprises at least one nuclease, which may be a ribonuclease, a deoxyribonuclease, or a nonspecific nuclease. The invention also relates to kits for the performance of various microbiological procedures, which kits comprise the nuclease inhibitors described herein.

The invention relates to 'a blood DNA preserving card and production method' and belongs to the biology technical field. A blood DNA preserving card comprises fiber-shaped media with the ability of absorbing DNA, and is characterized in that: the salinity is absorbed in the media and contains cell lysing reagent, nuclease inhibitor, a substance allowing the pathogenic bacterium and the virus protein to be denaturalized as well as anti-oxidization inhibitor. These salinities can inhibit the activities of the viruses to prevent the infection, can prevent the nuclease from gradating DNA, so that the blood DNA can be preserved indoor for a long-term. The pure DNA can be obtained from the preserved DNA in the card of the invention by bleaching the salt ions and blood corpuscle leftovers on the card media with the water, thereby avoiding the extraction process, ensuring the operation is simpler, being convenient to be operated in batches and saving the expense of the DNA extraction agent.

The invention discloses a preservative for preserving free DNA in peripheral blood. The preservative is composed of the following components in percentage by weight: 60 to 80% of ultrapure sterile water, 5 to 8% of anti-coagulant, 10 to 25% of antiseptic, 1 to 5% of metabolic inhibitor, and 0.5 to 3% of nuclease inhibitor. The provided preservative can preserve peripheral blood for at least 7 days at a normal temperature, 14 days at most, the blood can be transported at a normal temperature for 72 hours or more under the assistance of the preservative, 96 hours at most; and the free DNA amount is basically not affected during the period.

The invention discloses an animal nucleic acid sample normal temperature preservation reagent and an application thereof. The animal nucleic acid sample normal temperature preservation reagent comprises an acid cell lysis agent, nuclease inhibitors, a surfactant, sodium citrate and sodium acetate, wherein the acid cell lysis agent is guanidinium isothiocyanate, the used nuclease inhibitors are 8-hydroxyquinoline and beta-mercaptoethanol, and the used surfactant is lauryl sodium sulfate. On the other hand, the invention discloses a method for treating filter paper by using the animal nucleic acid sample normal temperature preservation reagent. The method comprises the following steps: 1) soaking the filter paper in the animal nucleic acid sample normal temperature preservation reagent; 2) baking the filter paper at 37-50 DEG C; 3) sterilizing the filter paper for later use.

The present invention concerns a compositions and method for isolating a nucleic acid from a cell-containing sample. There is disclosed a method comprising obtaining at least one cell-containing sample, which comprises a cell containing nucleic acid, obtaining at least one catabolic enzyme, obtaining at least one nuclease inhibitor, preparing an admixture of the sample, the catabolic enzyme, and the nuclease inhibitor, maintaining the admixture under conditions where the catabolic enzyme is active, and agitating the admixture, where the sample is digested to produce a nucleic acid-containing lysate of the sample.

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.

Methods and compositions for inhibiting and / or inactivating nucleases by employing nuclease inhibitors are provided. The nuclease inhibitors comprise anti-nuclease antibodies and non-antibody nuclease inhibitors. The anti-nuclease antibodies of the present invention may be a polyclonal or monoclonal antibodies, and may be anti-ribonuclease antibodies, anti-deoxyribonuclease antibodies, or antibodies to non-specific nucleases. A preferred embodiment comprises at least two nuclease inhibitors, and is referred to as a nuclease inhibitor cocktail. In some specific embodiments, the invention concerns methods of performing in vitro translation comprising obtaining a first nuclease inhibitor, which inhibitor is further defined as an anti-nuclease antibody, and placing the anti-nuclease antibody in an in vitro translation reaction. In many cases, the in vitro translation reaction comprises at least one nuclease, which may be a ribonuclease, a deoxyribonuclease, or a nonspecific nuclease. The invention also relates to kits for the performance of various microbiological procedures, which kits comprise the nuclease inhibitors described herein.

Mutant forms of ribonuclease inhibitor are described which are rendered more resistant to oxidation while retaining affinity for both ribonuclease and angiogenin. The mutant forms have another amino acid, typically an alanine, substituted for one or more of the adjacent cysteine residues in the wild-type sequence to prevent the formation of unwanted disulfide bonds which can disrupt the effectiveness of the molecule.

The invention provides a preserving fluid for biological sample nucleic acid detection and an application thereof. The preserving fluid is composed of a nuclease inhibitor, a surfactant, a chelating agent and a buffer solution. The preserving fluid is mixed with a solid sample or a liquid sample to denature proteins, inactivate pathogens, and inactivate DNA and RNA nuclease. The pathogens are inactivated, so the safety of a potential high-infectivity clinical sample is greatly improved, and good protection is formed for surrounding personnel and downstream operating personnel of the sample; the DNA and RNA nuclease is inactivated, so that the content and integrity of DNA and RNA in the sample are kept in a sampling state to the maximum extent, and the strict requirements on the aspects oftemperature, time and the like of clinical sample transportation and storage are reduced. The preserving fluid can be used for clinical high-infectivity nucleic acid detection of biological samples, and the biological samples comprise throat swabs, body fluids, lavage fluids, whole blood, whole blood cells, serum or plasma.

The invention relates to the field of virus nucleic acid detection. Specifically, the invention provides a pretreatment method for virus nucleic acid detection. The method comprises a step of mixing apretreatment solution containing a sample with a nucleic acid releasing agent and a qPCR amplification reagent. The pretreatment solution comprises Tris-HCl, EDTA-2Na, sodium chloride, a ribonucleaseinhibitor and an antibiotic and has a pH value of 6.5 to 8.0. A nucleic acid detection reaction solution prepared by the method can be directly used for qPCR, improves detection efficiency and shortens detection time.

The invention discloses a blood protection solution for preserving cfDNAs (cell-free DNAs) in whole blood at room temperature. The blood protection solution comprises anticoagulants, preservatives, nuclease inhibitors, metabolic inhibitors, cell stabilizers and membrane stabilizers. The blood protection solution has the advantages that by a blood cfDNA preservative, the nuclease activity can be inhibited, and the cfDNAs in the blood can be protected; white blood cells are stabilized, breakage of the white blood cells is prevented, and the cfDNAs are prevented from being contaminated by cell genomes; peripheral blood can be stored at the room temperature for 14 days and transported at the room temperature for at least 14 days, during which no obvious change occurs, so that the storage timeof blood samples at the room temperature is effectively prolonged, great assistance is brought to long-distance transport of the blood samples, the detection cost of the cfDNAs is saved, and application and promotion of cfDNA-related detection techniques are facilitated.

The invention relates to a protecting agent for free DNA in plasma and belongs to the biotechnical field. The protecting agent for free DNA in plasma is a sterile aqueous solution comprising the following components: a 50-600 g / L preservative, a 30-100 g / L nuclease inhibitor, a 20-200 g / L metabolic inhibitor and a 1-120 g / L anti-adsorbent. By mixing collected blood with the protecting agent in a proportion of (5-1000): 1, the free DNA of blood can be kept for 14 days in a constant temperature condition, the free DNA of blood can be kept for 4 days at 37 DEG C, and the free DNA of blood can betransported continuously for two days. By optimizing the components and the contents thereof, free nucleic acid in blood can be stored and extracted effectively, and meanwhile, nucleic acid doped intoblood cells as a result of splitting decomposition of hemocyte is avoided after blood is extracted.

The invention discloses a blood additive, which is prepared from 6.0 to 8.8 g / 100ml of anticoagulants, 2.0 to 8.4 g / 100ml of nuclease inhibiting agents, 4 to 20 g / 100ml of cell stabilizing agents, 0 to 0.31 g / 100ml of purine, 2 to 10 g / 100ml of metabolic inhibitors, 0.021 to 0.21 g / 100ml of apoptosis inhibitors and 0.1 to 1 g of nucleic acid protection agents. Non-Cl<-> buffer solution is 90 to 110 mM, and the pH is 7.3 to 7.5. The blood additive can be used for storing blood at the normal temperature for 14 days without influence on detection.

The invention provides nucleic acid preserving fluid. The nucleic acid preserving fluid consists of a nuclease inhibitor, a protein denaturant, a stabilizer, a bacteriostatic agent and a Tricine buffer solution, wherein the concentration of the nuclease inhibitor is 5mg / ml-20mg / ml, the concentration of the protein denaturant is 20mg / ml-80mg / ml, the concentration of the stabilizer is 30mg / ml-90mg / ml, the concentration of the bacteriostatic agent is 100mg / ml-500mg / ml, and the concentration of the Tricine buffer solution is 20nM-120nM. The nucleic acid preserving fluid has the beneficial effectsthat the nucleic acid preserving fluid is capable of effectively preventing degradation of nucleic acid at a normal temperature and providing a stable environment for nucleic acid, so that the preservation time of the nucleic acid is prolonged, and the safety during collection, transportation and detection can be guaranteed; and meanwhile, pathogenic microorganisms can be inactivated, and the nucleic acid preserving fluid has the advantages of low preparation cost, stable performance, easiness in preservation and the like.

The invention provides a cell preserving fluid, a preparation method and an application thereof and a cell preserving method and belongs to the technical field of cell preserving fluids. The cell preserving fluid comprises a cell protective agent, a nuclease inhibitor, a chelation stabilizer and a metabolism inhibitor. Under the synergistic effect of the four components including the cell protective agent, the nuclease inhibitor, the chelation stabilizer and the metabolism inhibitor, the obtained cell preserving fluid can effectively preserve CTC cells at the normal temperature for a long time, effectively protect the form and antigen state of the CTC cells, allow long-distance transportation and improve repeatability and reliability of CTC analysis and detection.

A blood anticoagulant used for protecting free DNA is disclosed. The blood anticoagulant comprises following components by a weight-to-volume ratio (W / V): 0.1-1.5% of a formaldehyde releasing agent, 1.5-2.2 g / L of an anticoagulant, 0.014-0.035 g / L of purine, 0.01-0.15% of dihydric phosphate, 0.4-1.0% of chloride salts and 0.3-1.4% of a nuclease inhibitor. The blood anticoagulant is a blood protecting agent effectively protecting cells from breaking, retarding degradation of free DNA in plasma, providing a stable environment for the plasma and prolonging blood stability, and avoids influences on subsequent detection which are caused by blood deterioration and DNA degradation.

The invention discloses a method for quickly extracting and preserving a silica gel membrane type DNA. By utilizing the characteristic of a silica gel membrane that the silica gel membrane adsorbs DNAunder the conditions of high salt and low pH and releases the DNA under the conditions of low salt and high pH, the invention does not need a process of extracting the DNA in advance after the silicagel membrane is processed by a novel solution containing a cell cracking agent and a nuclease suppressor, cells are directly lysed, the DNA is adsorbed on the silica gel membrane, the silica gel membrane adsorbing the DNA can be preserved for a long time because of the existence of the nuclease suppressor and the degradation of the DNA is avoided. In the extracting process, the pure DNA is adsorbed on the membrane by the elution function of eluent I after salt and cell residues on the membrane are rinsed. By the elution function of eluent II, the DNA combined on the membrane is eluted into the solution. The whole DNA extracting and preserving process has quick operation and low price and is simple and convenient.

The invention belongs to the technical field of biology, and particularly relates to a biological DNA (deoxyribonucleic acid) and RNA (ribonucleic acid) room-temperature preservation card and a manufacturing method thereof. The sample preservation medium of the preservation card is a material with water absorptivity or DNA combining capacity. The sample preservation medium contains a nuclease inhibitor and a nucleic acid stabilizer, and is subjected to DNase and RNase removal treatment. Compared with the prior art, the biological DNA and RNA room-temperature preservation card has the following advantages: the material with water absorptivity is adopted to preserve the dry DNA onto a film, and the film is subjected to enzyme removal treatment and treated by the nucleic acid stabilizer, so that the DNA and RNA can be preserved for a long time without degradation; the preservation card is free of toxic and harmful substances, and thus, is safe for the human and environment; all the used reagents and materials are free of PCR (polymerase chain reaction) and other enzymatic reaction inhibition components, and are suitable for the development of all downstream experiments in molecular biology; the preservation card does not need solid carbon dioxide in the transportation or mailing process, and can be transported at normal temperature; and the card information is detailed and is convenient for classified storage and search.

The invention provides a reagent composition for stabilizing nucleic acid molecules in a biological sample. The reagent composition comprises a pH stabilizer, a chelating agent, a protein denaturant, an antimicrobial agent and optional inorganic salt ions, a tissue penetrant, a reducer, a nuclease inhibitor and the like. The invention also provides a method for transporting, storing and stabilizing the nucleic acid molecules in the biological sample by using the reagent composition and application of the reagent composition in transporting, storing and stabilizing the nucleic acid molecules in the biological sample. The reagent composition ensures that high molecular weight DNA in the biological sample is kept for a long time at normal temperature.

The invention discloses a swab nucleic acid sample releaser and application thereof. The sample releaser comprises 1-20mol / L of metal ion chelating agent, 5-80mmol / L of buffer solution, 5%-40% (v / v) of polar organic solvent, 0.2%-10% (v / v) of surfactant and 0.1-1mol / L of nuclease inhibitor and has a pH value of 6-9. The sample releaser can integrate three steps, i.e., sampling preservation, deactivation and nucleic acid extraction into one step, and a sampled sample can be directly placed into a sample-releaser-containing sampling tube for preservation and transportation; and when in use, thesampling tube is mixed uniformly through up-down reversal, normal-temperature standing is carried out for 15 minutes after the sample and the sample releaser are uniformly mixed thoroughly, and then,nucleic acid of the sample can be released while deactivating the sample. Heating and mid-course uncapping are not required in a whole operating process from sampling to PCR detection, pollution to experiment operating personnel and laboratories is avoided effectively, and the operation is extremely simple; and in case of sudden infectious disease events, the sample releaser has the obvious advantages of rapidness, safety, stability, high efficiency, low cost and the like.

The invention discloses a water environment DNA collection and preservation card and a preparation method thereof, and relates to the technical field of biology. The problems that due to the lack of DNA separation and purification conditions at the collection site in existing conventional studies of water environment DNA, water samples need to be brought back to a laboratory for environmental sample processing and DNA extraction, storage and transportation condition requirements are high, the cost is increased, and the storage time is limited are aimed to be solved, the water environment DNA collection and preservation card includes a material with the ability to lyse cells and adsorb the DNA, the adsorption material is internally adsorbed with salinity, and the salinity includes a cell lysis agent, a nuclease inhibitor, a nucleic acid stabilizer and an antioxidant, wherein the cell lysis agent is one or more combinations of Tris hydrochloride, NP40 (ethyl phenyl polyethylene glycol),sodium chloride, guanidine isothiocyanate and SDS (sodium dodecyl sulfonate); and the nuclease inhibitor is a metal ion chelating agent, and the metal ion chelating agent is one of EDTA and an EDTA sodium salt.

The invention discloses a novel preservation solution for an alveolar lavage fluid microbial nucleic acid. The preservation solution comprises a treatment solution I and a treatment solution II, wherein the treatment solution I comprises a guanidine hydrochloride buffer solution, a nuclease inhibitor, a metabolism inhibitor, an alkali metal halide and glycerol, and the treatment solution II comprises an alkali metal hydroxide and a surfactant. The treatment solution I and the treatment solution II which are separately stored are used as the novel preservation solution for the alveolar lavage fluid microbial nucleic acid, so that the preservation solution can be prevented from generating precipitates, at the same time, different interference impurities can be treated respectively in a targeted manner, alveolar lavage fluid samples can be rapidly stabilized, the activity of nuclease is effectively inhibited, the cell lysis is prevented, at the same time, the free nucleic acid in the alveolar lavage fluid can be well protected, the preservation time is long, the accuracy and convenience of nucleic acid detection are improved, and thereby the preservation solution has great applicationvalue in the detection and diagnosis of tumor diseases, viruses and the like.

The invention provides a composition and method for preserving nucleic acid in urine. The composition provided by the invention contains the following components in percentage by weight: 2%-20% of a preservative, 1%-10% of a quenching agent, 2%-40% of a nuclease inhibitor, 5%-60% of a hydrophilic polymer, 1%-30% of a penetrating agent and 0.01%-10% of a metabolic inhibitor. The composition is capable of improving the stability of free nucleic acids in a biological sample and preventing release of nucleic acids in a karyocyte genome, and the separated free nucleic acids are widely suitable formass clinical diagnosis, development and application.

The invention provides a saliva preserving fluid and a preparation method thereof. The saliva preserving fluid comprises a cell stabilizer, a nuclease inhibitor, a surfactant, a protein denaturing agent and a preservative. The saliva preserving fluid has good broad-spectrum antibacterial activity, can inhibit pollution of bacteria, fungi and the like, enables genome DNA with good integrity and purity to still be obtained after a saliva sample has been preserved for one month at normal temperature, and can be applied to preservation of saliva of a subject in scientific researches such as gene detection and epidemiological investigation.

The invention belongs to the field of biological sample preservation, and discloses a virus sample direct amplification type sample preservation solution and an application method. The preservation solution comprises a surfactant, a buffer agent, salt ions, a reducing agent, alcohol and a nuclease inhibitor. The preservation solution provided by the invention can quickly inactivate viruses, realizes direct sample loading amplification detection, and avoids a nucleic acid extraction process with tedious steps.

The invention discloses a novel preservation solution for microbial nucleic acid in cerebrospinal fluid. The fluid comprises the following components of a nuclease inhibitor, a buffer solution, a protein denaturation inhibitor, trisodium citrate, a bacteriostatic agent and a stabilizer, and the pH value of the novel preservation solution for the microbial nucleic acid in the cerebrospinal fluid is7-8. According to the novel preservation solution for the microbial nucleic acid in the cerebrospinal fluid, the activity of nuclease can be rapidly and effectively inhibited, the nucleic acid in a cerebrospinal fluid sample is prevented from being degraded, the content of the nucleic acid in the cerebrospinal fluid is guaranteed, and meanwhile, the cell lysis is avoided, the detection backgroundnoise is reduced, and the accuracy of cerebrospinal fluid detection is improved; and the preservation fluid is relatively simple in components and low in cost, and meanwhile, the stability of the free nucleic acid can be maintained at room temperature, and the fluid is extremely suitable for wide clinical application.