39 results about "Guanidine thiocyanate" patented technology

The invention relates to the field of molecular biology, and discloses a faeces sample stabilizing solution. The stabilizing solution comprises the following components: 0.02M-4M Guanidine thiocyanate, Tris-HCI of which the pH is 5-10 and the concentration is 2-20 M, 0.05-0.1 mM NaCl and 1-15 mM EDTA. According to the invention, the stability of the detected components in the faeces sample can be effectively kept, self-help fetch by a patient at home can be possible, the faeces stabilizing solution cannot be a hindrance to downstream detection (nucleic acid extraction and detection), the mixture of the faeces and the stabilizing solution can directly enter a detection process, greatly facilitates the flexibility of the follow-up treatment process of the faeces sample, and is beneficial to the detection and the treatment of digestive system diseases.

The invention discloses a virus nucleic acid extraction kit based on nano magnetic beads. The lysis liquid comprises guanidine thiocyanate / guanidine hydrochloride, a nonionic surfactant, tris (hydroxymethyl) aminomethane hydrochloride and ethylenediamine tetraacetic acid, and the nonionic surfactant comprises one or more of triton X-100, Tween 20 and ethyl phenyl polyethylene glycol. The binding liquid is isopropanol, the volume ratio of isopropanol to pyrolysis is 1: 1, and the magnetic beads are silicon oxide hydroxyl nano magnetic beads. According to the kit, a nano magnetic bead extractionmethod is adopted, isopropanol is used as a binding liquid, making sample splitting, binding of nucleic acid and magnetic beads to be done in one step. The steps are simple, convenient and easy to implement, and the automatic extraction process of virus nucleic acid can be completed quickly at high flux within 9 min. The kit disclosed by the invention can be used for extracting RNA and DNA of viruses in sample, such as plasma, serum, nasopharynx swabs, urine, secretion liquid, virus concentrate, cell culture liquid supernatant and virus preservation liquid.

The invention provides a feces preservation solution, a preparation method thereof and a feces preservation method. The feces preservation solution contains 0.10-0.20 M of Tris base, 0.25-0.75 M of ethylenediaminetetraacetic acid (EDTA), 10 mM-room temperature saturated sodium chloride (NaCl), 5-15% of dimethyl sulfoxide (DMSO), 2.5-10% of ethanol and 5-20% of glycerol, wherein the pH of the fecespreservation solution is 7.0-9.0. The feces preservation solution can stably preserve microbial diversity in feces samples at normal temperature and improve DNA concentration, can ensure the accuracyof sequencing analysis of intestinal flora, realizes normal-temperature preservation and transportation of the feces samples, has a good preservation effect compared with an existing preservation solution, does not contain toxic components such as guanidine thiocyanate, and is safer to operate.

Disclosed is nucleic acid preserving compositions and methods of manufacturing and using the same. Compositions include a carrier, a chaotropic agent, a buffering agent, a chelating agent, a surfactant, an alcohol, an acid, and a mucolytic agent. Compositions as aqueous solutions can include water as a carrier. Preferred embodiments include water, guanidine thiocyanate, Tris, EDTA, SLS, SDA 3C, HCl, and N-acetyl-L-cysteine. Some embodiments include a colored dye as a visual indicator. Methods of manufacturing include combining the components into a mixture, such as an aqueous solution. Methods of use include providing a biological sample that includes nucleic acid and contacting the biological sample with the composition. Kits include the composition disposed in a portion of a biological sample collection apparatus.

The invention relates to a method for extracting genome and mitochondria DNA with a high salt method which is modified on the basis of the high salt method and the application thereof. Protein denaturant which contains guanidinium thiocyanate, guanidine hydrochloride and the like is used to crack cells in the method for extracting mitochondria DNA with the modified high salt method, which not only has the effect of depositing protein, but also has the effect of inhibiting nuclease, and the purity and the output of DNA can be greatly increased. The method can be faster and more convenient after being matched with an adsorption column, the operation of a single sample can be normally finished within 20 minutes, the high purity can be guaranteed through rinsing the column for many times, and the method can be directly used in PCR, Southern-blot and various endonuclease reactions. The method has convenient operation and low cost, and is in particular suitable for extracting the genome and the mitochondria DNA from clinical specimens (in particular in peripheral blood).

The invention discloses an extracting method for the total RNA (Ribonucleic Acid) of galangal. The method is improved on various aspects on the basis of the conventional guanidine thiocyanate-phenol-chloroform extraction method according to the characteristic of rich secondary metabolism product of the galangal and particularly rich polyhydric flavonol, i.e., acetone, water-soluble polyvinylpyrrolidone (PVP) and beta-mercaptoethanol are added in an RNA extracting solution. The acetone can be used for dissolving colored matters such as flavonols and the like; the water-soluble PVP can be sufficiently combined with phenol substances to form chelates; the beta-mercaptoethanol as a strong reducing agent can be used for providing a reducing condition so that polyphenol substances are not easy to oxidize; the three substances synergistically take effects, so that the interference and obstruction of the polyphenol substances are effectively inhibited. According to the invention, a crudely-extruded sediment of the RNA is treated by using Tris saturated phenol and high-concentration potassium acetate after being obtained, and part of proteins and polysaccharides can be simultaneously removed.

The invention relates to the technical field of a DNA extraction kit, in particular to a whole blood genome DNA extraction kit. The whole blood genome DNA extraction kit comprises lysate, an adjustingbead, a nano magnetic bead, a washing solution A, a washing solution B and an eluent; the lysate contains guanidine thiocyanate, sodium N-lauroyl sarcosinate, guanidine hydrochloride, proteinase k and Tween 20; the adjusting bead contains a semipermeable membrane balloon, and sodium chloride is put in the semipermeable membrane balloon; the nano magnetic bead is ferroferric oxide wrapping siliconoxide; the washing solution A contains hexadecyl trimethyl ammonium bromide, Tris-HCl, guanidine thiocyanate, absolute ethyl alcohol and FMES; the washing solution B contains Tris-HCl and absolute ethyl alcohol; the eluent contains Tris-HCl and disodium EDTA. When the kit is in use, the adjusting bead absorbs moisture produced in a fragmentation process and adjusts concentration of the lysate, the recovery efficiency of nucleic acid cannot be adversely affected, and accuracy of a detection result is guaranteed.

A formulation containing guanidine thiocyanate together with acetamide, one or more acetamide derivatives, or a combination of acetamide and one or more acetamide derivatives is used to purify one or more nucleic acids contained in a medium. In particular, a medium containing at least one nucleic acid is combined with a binding matrix and the formulation in order to cause the at least one nucleic acid to separate from its in vivo cellular environment and to bind to the binding matrix. The binding matrix with at least one nucleic acid bound thereto then is separated from substantially the rest of the combined medium and formulation, after which the at least one nucleic acid is eluted from the binding matrix to obtain the at least one nucleic acid in a substantially purified form. If different nucleic acids are to be selectively purified from a single medium, multiple binding matrices, each compatible with a different nucleic acid, can be used.

The invention discloses a method for extracting animal muscular tissue DNA easily, conveniently, safely and fast. The method comprises the steps that after pork, mutton, beef and chicken are washed with clean water, connective tissue and fat in animal tissue are removed; the animal tissue is shorn into small blocks of 200 mg, and the small blocks are placed in liquid nitrogen and fast ground till powder is obtained; 100 mg of the powder is taken to be placed in a 2-mL centrifuge tube, and the DNA is extracted through SDS splitting decomposition, isopropanol precipitating and adsorption column purification and can be directly used for PCR amplification. The DNA extracting method is adopted, and the whole process of DNA extraction of a sample can be completed within 2 hours. The quality standard of the extracted DNA meets the requirement of subsequent PCR amplification, meanwhile, the harm of various organic reagents to experiment staff in the extraction process of a traditional SDS method or guanidine thiocyanate method is avoided, and time is greatly shortened; the method is more suitable for achieving the aim of efficiently and fast extracting sample DNA in the animal source ingredient identification process, and has the advantages of being efficient, fast, low in cost, easy and convenient to operate, free of toxicity, safe and the like.

The invention discloses a kit for extracting dried blood spot genome DNA by a paramagnetic particle method and an extraction method. A DNA extraction reagent comprises a lysis solution, magnetic beads and a magnetic bead binding solution, wherein in the magnetic bead binding solution, the mass concentration of guanidine thiocyanate is 1-10 M / L, the mass concentration of Tris is 10-100 mM / L, the mass concentration of EDTA is 10-100 mM / L, and the volume fraction of Triton X-100 is 2%; or in the binding solution, the mass concentration of the guanidine thiocyanate is 1-10M / L, the mass concentration of the Tris is 10-100mM / L, the mass concentration of the EDTA is 10-100mM / L, and the volume fraction of the Tween 20 is 1.5%; and the magnetic beads are silicon hydroxyl magnetic beads. According to the dried blood spot genome nucleic acid kit and the extraction method disclosed by the invention, various blood-related samples, such as dried blood spots, blood samples, blood clot samples, blood gauze and tissue samples rich in blood cells, can be extracted; and the extraction efficiency is high, the purity is good, and the product stability is strong.

The invention discloses a magnetic bead method virus nucleic acid extraction kit and a use method, the magnetic bead method virus nucleic acid extraction kit comprises a lysis solution, a magnetic bead solution, a washing solution I, a washing solution II, a washing solution III and an eluent, the lysis solution is composed of the following substances: Tris-HCl, guanidine thiocyanate, EDTA, Triton-100, PEG-6000, SDS, isopropanol and water. The lysate provided by the invention has the beneficial effects that protease K does not need to be used, so that the cost is saved, heating lysis is generally needed during sample lysis, volatilization of an organic solvent can be accelerated, and the health of operators is harmed, but the lysate does not need to be heated, so that the harm to the operators can be reduced. The formula of the lysate for extracting nucleic acid is obtained by researching the proportion of the components of the lysate by the inventor.

The present invention relates to a method for detecting said nucleic acid while releasing the nucleic acid from a complex biological sample. The present invention relates to the use of cell lysis buffers containing strong chaotropic agents such as guanidinium thiocyanate to promote cell lysis and release of cellular nucleic acids. The invention also relates to the use of new forms of bicyclic nucleotide analogs to lock nucleic acids (LNA) Detection of specific nucleic acids released during cell lysis by nucleic acid hybridization. In particular, the covalent attachment method for capturing LNA-oligomers is described. New methods for sample preparation, such as polyadenylated mRNA, have also been proposed. The present invention further provides reagents for the method and applications of the method.

The invention relates to the technical field of biology, and particularly relates to a sperm DNA extraction method. The method comprises the following steps of: 1) taking a semen sample, cleaning thesemen sample, and performinglysis on the semen sample with a lysis buffer to obtain a lysate; 2) uniformly mixing the lysate with an alcohol solvent, and collecting a precipitate; and 3) washing the precipitate obtained in the step 2) with a sodium citrate solution, and collecting a precipitate, wherein the lysis buffer is prepared from 4.03-4.45M of guanidine thiocyanate, 0.095-0.105M of sodium chloride, 0.95-1.05% of sodium dodecyl sarcosinate, 0.1425-0.1575M of dithiothreitol and 0.19-0.21mg / mL of protease K, and is prepared by using sterile water. The sperm DNA extraction method provided by the invention is short in extraction time, large in total amount and high in quality, and can meet the requirements of various subsequent molecular genetic experiments.

The invention provides whole blood RNA quick lysate which does not need erythrocyte pyrolysis firstly, and an RNA extraction method. According to the method, the usage of toxic reagents is reduced toa large extent, RNA extraction steps are simplified, and RNA obtained through purification is good in integrity and high in purity and can be directly used for various molecular biology experiment ofRT-PCR, Northern blot, Dot blot and the like. Every 1000ml by volume of the whole blood RNA quick lysate disclosed by the invention consists of the following components by dosage of 0.2M-4M of guanidine thiocyanate, 0.2M-3M of ammonium thiocyanate, 0.01M-0.2M of anhydrous sodium acetate, 0.05M-0.3M of glacial acetic acid, 0.1%-2%(W / V) of sodium lauroyl sarcosine, 0.02M-0.3M of sodium chloride, 1%-15%(V / V) of glycerine, 10%-75% (V / V) of a water saturation phenol solution, and the balance DEPC treating water.

The invention discloses a fecal microorganism preserving fluid. The preserving fluid comprises a protein inhibitor, a stabilizer with a chelating agent and a preservative effect, a biological buffer solution, a metal chelating agent, sodium chloride and water, the final concentration of the protein inhibitor is 0.1-2 mol / L, the final concentration of the metal chelating agent is 1-100 mmol / L, thefinal concentration of the sodium chloride is 0.1-1 mol / L, the final concentration of the biological buffer solution is 1-100 mmol / L, and the final concentration of the stabilizer is 0.1% / L-10% / L. Theinvention further discloses a preparation method of the fecal microorganism preserving fluid, which comprises the follows several steps: step 1, adding 1 / 2 of sterile water by volume to a container;step 2, sequentially adding the sodium chloride, a Tris-CI buffer solution, ethylenediamine tetraacetic acid, the stabilizer and guanidine thiocyanate to the container; step 3, adding deionized waterto complete volume metering; and step 4, adjusting PH to 7.5, and performing sterilization and room-temperature cooling to obtain the preserving fluid. The preserving fluid has the advantages that thenormal-temperature preserving effect on samples is good, the preserving fluid is convenient to use, the whole preparation method is simple and the like.

The invention provides an excrement exfoliated cell nucleic acid preservation reagent as well as a preparation method and an application thereof. The excrement exfoliated cell nucleic acid preservation reagent comprises a nonionic surfactant, an anionic surfactant, an osmotic pressure regulator, a cell membrane stabilizer, a chelating agent, a pH buffer solution, water and a deodorant. The preservation reagent disclosed by the invention can be used for effectively cracking human exfoliated cells in excrement, and does not contain guanidinium thiocyanate, guanidinium isothiocyanate and other toxic components, so that a method for directly extracting and detecting the excrement genome from an excrement supernatant is provided, the interference of the excrement impurities can be effectively avoided, and the excrement sample does not need to be subjected to precipitation treatment before the detection.

The present invention discloses an extract and application thereof in RNA extraction. The extract includes, in terms of volume fraction, 30-65 vt% of water-saturated phenol, 9-15 vt% of glycerol and the balance of a solvent; an extract B also includes, in terms of molar concentration, 0.20-1 mol / L of guanidine thiocyanate, 0.4-0.8 mol / L of ammonium thiocyanate and 0.0073-0.018 mol / L of a surfactant; and the pH of the extract is 3.5-4.5. Components in the extract of the invention are reasonable in proportion design and have synergistic effects with one another, so that the purity of the extracted RNA is high, the yield is high, and the protein pollution is low; the extract are stable in performance, not easily affected by season, humidity and other conditions, and can be fully used in combination, so that the purpose of rapid, low-cost, high-quality and large-scale extraction of animal RNA in the laboratory can be realized.

The invention discloses a high-quality flora DNA extracting technique, which comprises four steps, namely sample collecting, preserving, pre-treatment and extracting, wherein in the step of extracting, firstly, a DNA self-sample is released from a lysis buffer solution containing cetyl trimethyl ammonium bromide; then, implementing separating and precipitating, so that a DNA precipitate is obtained; and finally, implementing adsorbing and purifying by virtue of guanidine thiocyanate and silicon dioxide, so that DNA, which is relatively high in purity, is obtained. The purity of the finished DNA prepared by the method is greater than 1.7 (OD260 / OD280), which is higher than that of a professional kit, and fragment is greater than 10kb, so that gene amplification and sequencing can be implemented. The method is simple and convenient to operate and easy for promotion and application; meanwhile, the method has the advantage of being good in universality; the high-quality DNA product can beobtained in various samples; and the method is applicable to related molecular experiment operations.

The invention discloses a detection kit for continuously monitoring lactic dehydrogenase isozyme I. The detection kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 is prepared from L-guanidine thiocyanate lactate biological buffer solution 1, preservatives and albumin bovine serum; the reagent R2 is prepared from oxidized coenzyme I potassium ferrocyanide biological buffer solution 2, and albumin bovine serum. The detection kit has the effects of high speed, sensitivity, high accuracy, high specificity and deterioration and volatilization difficulty.

The invention belongs to the field of molecular biological detection, and particularly belongs to the field of virus nucleic acid extraction. The invention discloses a composition for extracting 2019novel coronavirus nucleic acid. The composition is prepared from an extracting solution 1, an extracting solution 2 and an extracting solution 3, wherein the extracting solution 1 is prepared from guanidine hydrochloride, guanidine thiocyanate, urea, surfactin, potassium chloride, lithium chloride, triethanolamine lauryl sulfate, NP-40, triton X-100 and isopropanol; the extracting solution 2 comprises Tris-HCl, lithium chloride, sodium acetate, triethanolamine lauryl sulfate, NP-40, SDS and ethanol; and the extracting solution 3 comprises Tris-HCl, sodium chloride, and ethanol. By using the composition provided by the invention, the sensitivity of nucleic acid extraction and purification of 2019 novel coronavirus can be specifically improved, and the detection limit can reach 10 copies / mL.

The invention is related to diagnostic methods for detecting transmissible spongiform encephalopathies (TSEs) such as BSE and scrapie and related disease in humans. The invention provides use of guanidine thiocyanate (gdnSCN), or a functional equivalent thereof, for treating at least one sample derived from a mammal, including humans, for reducing the risk of scoring a false-positive test result in testing the sample for the presence or absence of aberrant prion protein.

The invention relates to a preserved lysate and a sample nucleic acid rapid extraction method based on the same. The preserved lysate comprises the following components in percentage by volume: 5-30% of absolute ethyl alcohol and 70-95% of a mixed aqueous solution, wherein based on the volume of the preserved lysate, the mixed aqueous solution comprises the following solutes in concentration: 0.5-1mol / L of ammonium chloride, 10-50mmol / L of trihydroxymethyl aminomethane hydrochloride, 0.4-2mol / L of guanidine thiocyanate, 0.4-1mol / L of sodium chloride, 10-50mmol / L of ethylenediamine tetraacetic acid and 0.1-2mmol / L of tris- (2-formylethyl) phosphine hydrochloride. The method comprises the following steps of: (a) putting a sample into a closed container filled with nano beads and the preserved lysate; (b) lysing the sample; and (c) incubating at room temperature, and performing centrifugal washing. By virtue of the prepared reagents, full lysis of cells and nucleic acid protection are realized by combining chemical and physical methods.

The invention provides an RNA purification liquid as well as a preparation method and application thereof. In the RNA purification liquid, the purification liquid contains at least the following components based on the total amount of the purification liquid: sodium dodecyl sulfate with a content of 5 g / 100ml-50 g / 100ml, guanidine thiocyanate with a content of 10 g / 100ml-30 g / 100ml, sodium citratewith a content of 0.01 g / 100ml-2 g / 100ml, N-alkanoylsarcosine with a content of 0.01 g / 100ml-2 g / 100ml, mercaptoethanol with a volume percentage of 0.01%-2%, polyethylene glycol octylphenyl ether with a volume percentage of 0.01%-2%, ethylenediaminetetraacetic acid disodium salt with a content of 0.01 g / 100ml-0.5 g / 100ml, and solvent water. The RNA purification liquid provided by the invention has low costs and a good purification effect, and a purification method is simple and rapid, can be completed within 40 minutes, and can be carried out under room-temperature conditions.

A process for extracting RNA from cotton tissue includes such steps as extracting in guanidine thiocyanate-chloroform solution, dissolving the coarse extract in the cracking liquid containing the polyvinyl pyrrolidone 4000 and beta-mercaptoethanol, and purifying by water-saturated phenol.

The invention provides a reagent kit for influenza a H3N2 type fluorescent PCR diagnosis. The reagent kit comprises RNA extraction solutions and a PCR reaction solution. The RNA extraction solutions comprise the RNA extraction solution I, the RNA extraction solution II, the RNA extraction solution III and the RNA extraction solution IV. The RNA extraction solution I is prepared from lauryl sodium sulfate, triton, guanidine thiocyanate and magnetic beads. The RNA extraction solution II is prepared from 4-(2-hydroxyerhyl) piperazine-1-erhaesulfonic acid and sodium chloride. The RNA extraction solution III is prepared from triton and sodium chloride. The RNA extraction solution IV is prepared from mineral oil. The PCR reaction solution comprises an upstream primer, a downstream primer and a probe, wherein the upstream primer and the downstream primer are used for target polynucleotide amplification, and the probe is used for target polynucleotide detection. The reagent kit is an influenza a H3N2 type fluorescent PCR diagnosis product excellent in RNA extraction rate and detection sensitivity. Typing detection can be carried out on influenza a viruses in throat swabs and various samples to judge whether patients are infected with the influenza a H3N2 type or not and assist doctors in making better diagnosis and more precise treatment.