21 results about "Proteinase K" patented technology

The invention provides an in-vitro induction and purification method for Giardia lamblia cysts. The induction method is characterized in that Giardia lamblia trophozoite is cultured in a TYI-S-33 culture medium containing pulvis fellis suis to obtain the Giardia lamblia cysts. According to the in-vitro induction method for the Giardia lancea cysts, the pig gall powder is added into the TYI-S-33 culture medium to induce the Giardia lancea trophozoite, and it is found that compared with common ox gall powder, the proportion of converting the Giardia lancea trophozoite into the cysts can be remarkably increased, and the generation rate of the induced cysts stably reaches 50%; according to the giardia lamblia cyst purification method disclosed by the invention, protease K is utilized for purification, so that pure cysts can be obtained, long-term stability and activity can be maintained, and the cysts can be stably stored for several weeks to several months at 2-8 DEG C and can be prepared into giardia lamblia cyst standard substances; the method is used for method verification, quality control, disinfection efficiency research, filtration efficiency evaluation and the like.

This invention relates to a method and composition for dispersing biological tissues with the aim of obtaining single cells for analysis. More specifically, the composition of the present invention is an enzyme composition comprising proteinase K. The composition of the present invention can be used in a method for effectively dispersing tissues at low temperatures for applications in high-throughput single-cell omics research.

Capture mixtures and activated capture mixtures are provided that are useful for nucleic acid separation and purification are provided. The mixtures comprise lithium lauryl sulfate, lithium hydroxide, a zwitterionic sulfonic acid buffering agent, and optionally, proteinase K, capture probes comprising a first specific binding partner (SBP), and a second specific binding partner immobilized to a solid support. Related combinations, methods, uses, and kits, are also provided.

The application discloses a DNA / RNA co-extraction method and kit based on a double-water-phase temperature-induced phase separation, which utilizes polyethylene glycol-sodium citrate to form a primary double-water-phase system at 18-25 DEG C, and by adding an ionic liquid modified temperature-sensitive polymer (IL-NIPAM) with a LCST of 32 DEG C plus or minus 1 DEG C, the IL-NIPAM selectively binds with RNA, and the separation of RNA and DNA in a biological sample is realized by temperature regulation. The DNA / RNA co-extraction kit includes a double-water-phase premix, and the IL-NIPAM is added in the double-water-phase premix in advance or during use, and the use concentration of the IL-NIPAM is 0.5-1.5% w / w. The method does not need proteinase K digestion and does not need to add organic solvents, and can quickly and efficiently recover DNA, mRNA and miRNA, and is suitable for the co-extraction of nucleic acids from trace samples such as fingertip blood, dry blood spots and FFPE sections, and is suitable for automation and large-scale production.

This invention relates to the field of nucleic acid release, specifically disclosing a rapid nucleic acid release reagent, kit, method, and application for animal tissue genotyping. The release reagent comprises a weakly alkaline reaction buffer, an enzyme-catalyzed reaction enhancer, a nonionic surfactant, a nucleic acid amplification enhancer, a first component composed of histidine, trehalose, and glutathione in a specific ratio, and a second component with a visual indicator function; a matching thermosensitive proteinase K formation kit is also included. The kit can be pre-packaged in multi-well reaction plates for improved ease of use. The release method of this invention achieves rapid release of nucleic acids from animal tissues through pre-incubation activation, mild-temperature lysis, and high-temperature enzyme inactivation steps. Simultaneously, the first component exhibits a strong synergistic effect with the thermosensitive proteinase K, effectively protecting the integrity of the nucleic acids.

The invention relates to a preparation method of a narrow gadus bone peptide compound and chelated calcium of the narrow gadus bone peptide compound. The method comprises the following steps: by taking a walleye cod processing byproduct fish bone as a raw material, carrying out trypsin preliminary enzymolysis, ultrafiltration and protease K targeted enzymolysis to obtain the walleye cod bone peptide compound. The narrow gadus bone peptide compound comprises one or more active peptides of DFF, DIF, DNAF, DPF and DYF. Chelated calcium can be prepared by using the narrow gadus bone peptide compound or the narrow gadus bone active peptide. Waste resources are fully utilized, high value of low-value products is achieved, environmental pollution is reduced, high economic and social benefits are achieved, and the prepared chelated calcium with the high additional value can be applied to the fields of dietary nutrition, biological medicine products and the like.

This invention discloses a preservation solution for preserving sample DNA at room temperature and its preparation method, belonging to the field of biological sample preservation technology. The preservation solution comprises: a lysis system, a nucleic acid protection system, and a buffering and stabilizing system; the lysis system comprises a complex surfactant and an enzymatic hydrolysis aid; the complex surfactant comprises: polyether polyol fatty acid ester and cocamidopropyl hydroxysulfonate betaine; the enzymatic hydrolysis aid comprises lysozyme Lyso-V and proteinase K; the nucleic acid protection system comprises: nitrogen-heterocyclic polyamine-carboxylic acid derivatives and dextran sulfate; the nitrogen-heterocyclic polyamine-carboxylic acid derivatives comprise: 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''tetraacetic acid, 1,4,7-triazacyclononane-N,N',N''-triacetic acid, disodium ethylenediaminetetraacetate-nitrocyclic derivative, and diethylenetriaminepentaacetic acid-piperazine derivative; the buffering and stabilizing system comprises: an amphoteric buffer and a polymeric stabilizer; the amphoteric buffer comprises: 2-(N-morpholino)ethanesulfonic acid and N-tris(hydroxymethyl)methylglycine.

This invention belongs to the field of biotechnology and discloses a plasma cell-free DNA extraction kit, extraction method, and applications. The kit includes magnetic beads, lysis buffer, proteinase K, binding buffer, washing buffer, and elution buffer. The extraction method involves mixing the plasma sample, lysis buffer, proteinase K, binding buffer, and magnetic beads. The lysed plasma cell-free DNA (cfDNA) is incubated with the magnetic beads to form a magnetic bead-cfDNA complex. Under a magnetic field, the complex is washed to remove various impurities, and the plasma cfDNA is obtained after elution. The method of this invention is simple and rapid, with short sample lysis time and stable quality. Replacing isopropanol with anhydrous ethanol during lysis results in high nucleic acid recovery, making it suitable for plasma cfDNA extraction. The purity is suitable for subsequent sequencing and PCR / Q-PCR. The magnetic bead method is beneficial for large-scale use and has significant implications for research and clinical applications.

The application provides a method for improving the detection rate of norovirus in aquatic products, which comprises the following steps: first, using a proteinase K solution to enzymolysis the digestive gland of shellfish, then adding a TGBE solution to elute, then adding enzymolysis corn starch to block activated carbon to remove the RT-qPCR inhibitor in the detection sample, taking the supernatant to extract nucleic acid, and using a fluorescent quantitative PCR instrument to detect. The detection method of the application reduces the inhibitor in the shellfish sample which interferes with the fluorescent quantitative PCR by using enzymolysis corn starch. In combination with the proteinase K enzymolysis, the TGBE elution and the RT-qRCR, the norovirus in the shellfish sample is detected, and the detection rate of the shellfish sample is increased by 10-20 times in 4h compared with the national standard method. The virus recovery rate and the detection sensitivity are greatly improved, and the method is suitable for shellfish samples, and has the characteristics of simple operation, high detection sensitivity and short time consumption.

The invention belongs to the technical field of molecular biology and in-vitro diagnosis, and particularly discloses a lysis solution composition and application thereof in preparation of a reagent for extracting genome DNA. The lysate composition comprises the following components: a pH regulator, 0.05-1.0% (v / v) of a nonionic surfactant, 0.1-2 mg / mL of bovine serum albumin, 0.5-5 mM of a reducing agent and 50-500 g / mL of protease K, and the pH value of the lysate composition is 7.0-8.0. The invention provides the genome DNA quick release lysate which is reasonable in composition and simple and convenient to operate, efficient release and stable storage of DNA in a complex biological sample are realized under the condition that a traditional purification step is not needed, and the obtained DNA can be directly used for nucleic acid amplification detection such as PCR (Polymerase Chain Reaction) and LAMP (Loop-Mediated Isothermal Amplification), so that the detection efficiency, repeatability and application convenience are improved; the actual requirements of rapid molecular diagnosis and on-site detection are met.

This invention provides a nucleic acid extraction and release agent for the detection of multiple pathogenic microorganisms, comprising the following components: Tween 20, proteinase K, EDTA, and KOH. The rapid nucleic acid extraction and release agent of this invention, after incubation with the sample, completes the nucleic acid extraction and release process, improving extraction efficiency, reducing reagent and sample usage, simplifying operation steps, and lowering labor costs. It has advantages such as high efficiency, low cost, and high quality.

This invention belongs to the field of food processing technology, specifically relating to an oat bran β-glucan extract, its preparation method, and its application. The preparation method involves obtaining the oat bran β-glucan extract from oat bran through multi-stage enzymatic hydrolysis. The enzymes used in the multi-stage hydrolysis sequentially include pentosanase, thermostable α-amylase, and proteinase K. This invention, under ultrasound assistance, allows pentosanase to efficiently break down cell walls first, significantly increasing the release rate of β-glucan. Subsequently, thermostable α-amylase and proteinase K precisely remove starch and protein impurities sequentially, greatly improving the purity of the product. This process route is mild, highly specific, and operates continuously, reducing impurity residue while improving extraction efficiency, providing a reliable and cost-effective solution for the large-scale preparation of high-purity, highly bioactive oat β-glucan.

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 relates to the technical field of cell culture and biological materials, and discloses a morphology-adjustable three-dimensional cell culture scaffold and a preparation method and application thereof.The preparation method comprises the following steps that oligopeptide Fmoc-FF is dissolved in an organic solvent to prepare an Fmoc-FF solution, the Fmoc-FF solution and a protease K aqueous solution are mixed, and a three-dimensional cell culture scaffold is obtained; the self-assembly process of the Fmoc-FF is initiated, and hydrogel is formed; after the Fmoc-FF forms hydrogel, protease K in a system is removed by utilizing the shear thinning characteristic of the hydrogel, and the Fmoc-FF nanofiber which is free of protease K residues and has a specific microstructure is obtained. According to the method, the diameter and length of the self-assembled nanofiber are adjusted by changing the adding concentration of protease K under the enzymatic hydrolysis effect of protease K on Fmoc-FF dipeptide. The method avoids the problem of high coupling of micro-morphology and macro-mechanical properties in a traditional collagen hydrogel system.

The present application relates to the field of biotechnology, in particular to a blood cell pathogenic microorganism host nucleic acid extraction kit and method, comprising cell lysis solution, DNA enzyme, DNA enzyme buffer, proteinase K, lysozyme, cell wall lyase, glass beads and extraction lysis solution;The cell lysis solution is saponin and beta octyl thioglucoside solution;The DNA enzyme is Ultra Nuclease;The DNA enzyme buffer comprises MgCl2, Tris-HCl and BSA;Low concentration of saponin and low concentration of beta octyl thioglucoside are used, the detection of various pathogenic microorganisms is considered under the condition of ensuring good host removal effect. The chemical method and physical method are combined to break the wall of microbial cells, the positive detection rate of pathogenic microorganisms in clinic is improved, and the blank of pathogenic microorganism missed detection caused by the detection of plasma sample cfDNA at present is made up.

The application discloses an extractable nucleic acid trichomonad activity protection solution and relates to the technical field of biotechnology, which comprises two component reagents A and B, the reagent A comprises 0.8-1.0% NaCl, 0.1-5% Tris-Cl buffer, 0.1-1.5% SDS, 1.5-5% glucose, 5-15% serum, 0.5-5% betaine, and the balance is sterile deionized water, and the reagent B is a proteinase K solution. The application can not only dilute secretion, preserve trichomonas and culture trichomonas, but also be used for extracting trichomonad nucleic acid in PCR molecular experiment, so that the application can realize double application of trichomonad detection. The application not only helps to solve the health problems of women and improve fertility protection, but also is the first domestic invention to provide an example for new technology transformation service to the society.

The invention discloses a method, a kit and a server for identifying species of a gentian medicinal material in a mixture. According to the invention, the optimized lysate formula significantly improves the quality of DNA extracted from a complex mixture. The reducibility of the lysate is enhanced by increasing the content of beta-mercaptoethanol, and the DNA is effectively prevented from being oxidized and damaged in the extraction process; the added protease K strengthens the digestive ability to protein, and reduces the wrapping and pollution of protein impurities to DNA (Deoxyribose Nucleic Acid); due to the addition of PVP, polyphenol substances in the mixture can be adsorbed, and degradation caused by combination of the polyphenol substances and DNA is avoided; the lysate can better cope with various interference components in a complex matrix due to the moderately improved CTAB concentration and salt concentration and stronger metal ion chelating ability, and meanwhile, the pH value of the solution is stabilized by the adjusted Tris-HCl concentration, so that the DNA is kept stable in the extraction process. Due to the improvements, the purity and the integrity of the extracted DNA are obviously improved, and a high-quality template is provided for subsequent species identification.

The invention belongs to the technical field of port animal quarantine, and discloses an imported animal skin pathogenic microorganism rapid early warning method based on metagenome sequencing, and the method comprises the following steps: optimization pretreatment of an imported animal skin sample: taking the imported animal skin sample, carrying out sterile grinding to prepare a homogenate, and carrying out spray drying on the homogenate; carrying out nucleic acid enrichment by combining protease K + lysozyme composite enzymolysis with a magnetic bead method; carrying out metagenome sequencing and bioinformatics analysis; performing biological safety risk research, judgment and early warning: performing risk grading on pathogens and generating early warning information on the basis of an analysis result in the step S2 and in combination with an imported non-edible animal product monitoring technical table; efficient and accurate detection of target pathogens: aiming at the high-risk pathogens early-warned in the step S3, developing a standardized detection reagent by adopting at least one technology of CRISPR / Cas, recombinant RPA and digital PCR, and carrying out targeted verification detection on the sample. The sample pretreatment is adaptive to the characteristics of animal skin, so that the enrichment efficiency of pathogenic nucleic acid is improved; a multi-technology detection system is combined, and the requirements of'broad-spectrum early warning 'and'accurate verification' are considered; technical support is provided for port animal quarantine, and biological safety hazards are effectively prevented and controlled.

The invention belongs to the technical field of biology, and particularly relates to a biological matrix DNA extraction method which comprises the following steps: 1) taking a biological matrix, adding a solvent for homogenizing, adding a protease K solution for cracking and centrifuging, and taking a supernatant to obtain a biological sample solution; and 2) taking the biological sample solution, the magnetic beads and the eluent obtained in the step 1), and performing nucleic acid extraction in a full-automatic nucleic acid extraction and purification instrument. The extraction method provided by the invention has high extraction efficiency on tissue samples (heart, liver, lung, kidney, brain, muscle, stomach, testis, duodenum and uterus) and secretions (excrement), and can be used for accurate PCR detection.

The invention discloses a method for capturing an FFPE sample space transcriptome and application of the method. The method comprises the following steps: exposing mRNA on an FFPE slice by using a de-crosslinking reagent, transferring RNA to a space transcriptome capture chip, carrying out reverse transcription, melting, synthesizing two chains, carrying out PCR (Polymerase Chain Reaction) amplification enrichment and purification, constructing a library, and carrying out sequencing analysis. The decrosslinking reagent comprises any one of the following components: (1) a NaCl solution; (2) a PBS buffer solution containing urea and Tween; (3) preparing a protease K solution; and (4) a sodium citrate buffer solution. Compared with the traditional method, the method disclosed by the invention has the advantages that the integrity of RNA (Ribonucleic Acid) can be better reserved, the RNA with polyA which is not completely degraded is efficiently enriched, and finally, the space transcriptome capture analysis of a low-quality FFPE sample is realized.