10 results about "Guanidine isothiocyanate" patented technology

This invention relates to the field of fecal genomics kit technology, and particularly to a safe, rapid, and high-throughput kit for extracting fecal genomes and its usage method. The kit includes a sample preservation solution, magnetic beads, a lysis buffer, a washing buffer 1, a washing buffer 2, and an elution buffer. The sample preservation solution is composed of guanidine isothiocyanate, sodium citrate, Triton X-100, Tween 20, propidium iodide, and purified water. The magnetic beads are hydroxyl magnetic beads. The lysis buffer is composed of guanidine hydrochloride, Tris base, Tris hydrochloric acid, Triton X-100, isopropanol, and purified water. Washing buffer 1 is composed of guanidine hydrochloride solution and isopropanol. Washing buffer 2 is composed of anhydrous ethanol and purified water. This kit can easily extract the genome from fecal samples, is simple to operate, safe and non-toxic, and can extract multiple samples at once, enabling rapid diagnosis and filling the gap in the current market for automated high-throughput kits for extracting genomes from fecal samples.

The present application relates to the technical field of nucleic acid extraction and purification, and particularly relates to a DNA and RNA co-extraction reaction reagent and an extraction method, the reaction reagent comprising a nucleic acid protective agent RP, a sample lysis and binding liquid LB, a protein washing liquid WB1, a nucleic acid rinsing liquid WB2, and a nucleic acid elution liquid EB; the nucleic acid protective agent RP comprising DTT, mercaptoethanol, TCEP, and sodium hydroxide; the sample lysis and binding liquid LB comprising Tris-HCl, benzenesulfonic acid, sodium citrate, guanidine hydrochloride, guanidine isothiocyanate, guanidine thiocyanate, urea, triton X-100, SDS, potassium chloride, sodium chloride, bromhexine, N-acetylcysteine, ambroxol hydrochloride, isopropyl alcohol, and anhydrous ethanol; the protein washing liquid WB1 comprising Tris-HCl, benzenesulfonic acid, sodium citrate, guanidine hydrochloride, guanidine isothiocyanate, guanidine thiocyanate, urea, triton X-100, sodium chloride, isopropyl alcohol, and anhydrous ethanol; the nucleic acid rinsing liquid WB2 comprising Tris-HCl, sodium chloride, isopropyl alcohol, and anhydrous ethanol; and the nucleic acid elution liquid EB being nuclease-free water; the reagent can rapidly and fully enrich and purify DNA and RNA of a sample simultaneously.

The invention relates to a kit for extracting DNA from FFPE tissue. The kit comprises a dewaxing solution, a lysis solution and a denaturation solution, the dewaxing liquid contains mineral oil, dimethyl silicone oil and butyl laurate; the lysis solution contains sucrose, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride, ethylenediamine tetraacetic acid, polysorbate-20 and water; the denaturation liquid is prepared from guanidine isothiocyanate, trihydroxymethyl aminomethane salt, sodium chloride, ethyl phenyl polyethylene glycol, triton X-100, lauryl sodium sulfate and water. According to the kit, the use of a traditional toxic organic reagent dimethylbenzene is avoided, and dewaxing liquid, lysate, denaturation liquid and an extraction method are improved, so that the nucleic acid fragmentation is reduced, the nucleic acid yield is increased, the genome DNA can be safely, quickly and efficiently extracted from an FFPE sample, the yield and integrity of the DNA are improved, and the kit is suitable for downstream DNA sequencing analysis.

This invention relates to the field of biological sample preservation technology, specifically to a room-temperature preservation solution for bacterial RNA from sputum samples of tuberculosis patients. Each 100 mL of the RNA preservation solution comprises the following ingredients: 2-5 g tris(hydroxymethyl)aminomethane, 0.5-1.3 g mucin digester, 20-30 g guanidine isothiocyanate, 0.1-0.3 g hexadecylpyridine chloride, 0.5-1.2 g ethylhexene dicysteine, 20-40 mL fixative, 1-5 g stabilizer, 0.1-0.5 g inorganic salt, 0.3-0.6 g surfactant, and 0.1-0.5 g preservative. The preservation solution provided by this invention can effectively liquefy sputum samples at room temperature, meeting the requirements for long-distance sample transportation. The RNA quality of sputum samples treated with the preservation solution of this invention is suitable for various subsequent molecular detection methods, possessing significant application value and promising prospects for widespread application.

The application relates to a sample processing method for cervical exfoliative cell PCR detection, which adopts a sample processing reagent, including a processing liquid, a protection liquid, a conversion liquid, a first purification liquid, a second purification liquid, a third purification liquid and a collection liquid. The processing liquid contains 0.1-0.5N sodium hydroxide and 4-8mol / L urea; the protection liquid contains 1-8mM guanidine isothiocyanate, 0.1-5% SDS, 5-150mM Tris-Hcl and 5-100mM EDTA; and the conversion liquid contains 100-500mM sodium bisulfite, 50-250mM ammonium sulfite, 2-15M ammonium bisulfite, 0.1-3M tetrahydrofurfuryl alcohol, 1-5mM TEPA and 0.1-2M magnesium sulfite. The sample is lysed and protected by the processing liquid and the protection liquid, direct conversion and detection are facilitated, processing time and steps are saved, DNA loss is reduced, cost is saved, the accuracy and reliability of detection results are ensured.

This invention provides a nucleic acid extraction reagent, kit, and method. The nucleic acid extraction reagent includes a pretreatment solution, proteinase K, lysis binding buffer, magnetic beads, washing buffer 1, washing buffer 2, washing buffer 3, and elution buffer, all stored separately but used in combination. The pretreatment solution has a pH of 7.80-8.20 and is composed of 0.1%-1% sodium dodecyl sulfate, Tris-HCl solution, and EDTA-Na2 solution by mass. The lysis binding buffer has a pH of 7.80-8.20 and is composed of 3M-4M guanidine isothiocyanate, 11%-15% polyethylene glycol 6000 by mass, 5%-10% Triton X-100 by mass, Tris-HCl solution, and EDTA-Na2 solution. This invention achieves "high concentration, high purity, and high efficiency" nucleic acid extraction through innovative extraction reagent components (non-toxic lysis system), reagent kit design (high-throughput pre-packaging), method optimization (automated coupling), and application expansion (adaptation to frozen samples), providing a reliable template for molecular diagnostics.

The application provides an alcohol-free and proteinase K-free nucleic acid extraction reagent and a nucleic acid rapid extraction method, and relates to the technical field of molecules.The extraction reagent comprises a lysis solution, and the lysis solution comprises guanidine isothiocyanate with a molar concentration of 2-4 M, a thiol reducing agent with a volume fraction of 1%-5% v / v, sodium erythorbate with a mass fraction of 0.02%-0.5% w / v and a surfactant with a mass fraction of 1%-10% w / v; and the lysis solution has a pH of 7-8.5.The lysis solution can be stably stored at room temperature for a long time; in application, only one washing solution is matched, one-step washing is performed, the complexity of nucleic acid extraction can be reduced, and the extraction time is shortened; and the lysis solution does not contain alcohol reagents, and there is no need to dry before nucleic acid elution.The technical problems that, in the prior art, the lysis solution has complex components, proteinase K needs to be matched, many impurities are introduced, multiple washing is needed, the steps are complicated, and a long time is consumed are solved.

The invention provides an extraction reagent for extracting small RNA (Ribonucleic Acid) in circulating body fluid. The extraction reagent comprises a splitting combination solution containing Tris-HCl, SDS (Sodium Dodecyl Sulfate), EDTA (Ethylene Diamine Tetraacetic Acid), NaCl, sodium citrate and guanidine isothiocyanate, the invention also provides a kit containing the extraction reagent, an extraction method and a library building method of small RNA in body fluid. According to the method, a set of special extraction process is established for extracting extremely trace small RNA in body fluid, and direct and efficient extraction of small RNA in circulating body fluid is realized by optimizing the components and the proportion of the cracking combination liquid and combining a conventional magnetic bead purification technology; in the library construction process, high-quality on-machine sequencing of the final library is realized by finely adjusting the proportion of the linker addition amount in each step and optimizing library extraction. According to the high-throughput sequencing method disclosed by the invention, the initial dosage of the sample is reduced while the operation process is simplified, and the success rate of library construction and the overall quality of the sequencing library are remarkably improved.

The invention relates to the technical field of biological sample preserving fluid, in particular to normal-temperature preserving fluid for bacterial RNA (Ribonucleic Acid) of a sputum sample of a tuberculosis patient. Every 100 mL of the RNA preserving fluid is prepared from the following raw materials: 2 to 5 g of tris (hydroxymethyl) aminomethane, 0.5 to 1.3 g of a mucus digesting agent, 20 to 30 g of guanidine isothiocyanate, 0.1 to 0.3 g of cetylpyridinium chloride, 0.5 to 1.2 g of ethylidene dicysteine, 20 to 40 mL of a fixing agent, 1 to 5 g of a stabilizing agent, 0.1 to 0.5 g of inorganic salt, 0.3 to 0.6 g of a surfactant and 0.1 to 0.5 g of a preservative. The preserving fluid provided by the invention can effectively liquefy a sputum sample under a normal temperature condition, and meets the long-distance transportation requirement of the sample. The RNA quality of a sputum sample treated by the preserving fluid is suitable for a plurality of subsequent molecule detection methods, and the preserving fluid has important application value and popularization prospect.

The invention belongs to the technical field of nucleic acid detection, and particularly relates to a tongue swab sample preserving fluid and application thereof. Comprising 20 mM to 100 mM of Tris-HCl or HEPES, 0.3% to 0.6% (v / v) of Tween 20, 0.04% to 0.08% (v / v) of Triton X-100, 0.03% to 0.05% (w / v) of guanidine isothiocyanate, 0.1 mM to 0.3 mM of EDTA-2Na (Ethylene Diamine Tetraacetic Acid), 0.1 wt% to 0.2 wt% of cane sugar and 20 mM to 25 mM of NaOH. The pH value of the tongue swab sample preserving fluid is 7-8, and the tongue swab sample preserving fluid can adapt to enriched magnetic beads, so that effective extraction of low-abundance mycobacterium tuberculosis nucleic acid in a tongue swab sample is realized.