21 results about "Proteinase C" 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.

The invention discloses a kit and a method for extracting nucleic acid from an ophthalmic sample and application of the kit and the method in metagenome sequencing. A method of extracting nucleic acid from an ophthalmic sample includes extracting nucleic acid from the ophthalmic sample using a sample processing system comprising the ophthalmic sample, a carrier RNA, protease K, and a lysis reagent. According to the method, a carrier RNA is added to prevent loss of trace nucleic acid, high-concentration protease K is adopted to enhance digestion of protein, and a chemical reagent is adopted to lyse cells, so that excessive damage and degradation of nucleic acid and low-volume elution caused by severe oscillation of mechanical wall breaking are avoided, and the final concentration of nucleic acid is improved.

The application discloses a Cas protein PAM analysis method based on local geometric structure and machine learning, and belongs to the fields of biotechnology and artificial intelligence. The method comprises the following steps: constructing an initial structure of a Cas protein-DNA-RNA ternary complex by using a structure prediction tool, and generating a mutant structure data set containing different PAM sequences by calculation and optimization; extracting and embedding the local structure of a PAM site to obtain a structure feature vector, performing dimension reduction and cluster analysis by using an unsupervised machine learning method, realizing natural classification of the PAM sequence, and identifying the cluster result in combination with P prior biological knowledge. The application creatively combines the three-dimensional local geometric structure of a protein and machine learning, can predict and classify the PAM preference of a Cas protein from a structure source, has the advantages of objectivity, comprehensiveness, and the ability to reveal hidden patterns, and can be verified by experimental data.

The present invention relates to methods and compositions for dissociating biological tissue with the aim of obtaining single cells for analysis. More specifically, the compositions of the invention are enzyme compositions comprising protease K. The compositions of the invention are useful in methods for efficiently dissociating tissue at lower temperatures for application in high throughput unicellular omics studies.

The invention relates to the field of astaxanthin synthesis, and discloses recombinant saccharomyces cerevisiae as well as a construction method and application thereof. The recombinant saccharomyces cerevisiae provided by the invention comprises the following exogenous genes: coding genes of beta-carotene hydroxylase CrtZ, beta-carotene ketolase CrtW, nicotinamide adenine dinucleotide kinase, ferredoxin-NADPH (nicotinamide adenine dinucleotide phosphate) oxidoreductase and ferredoxin; on the basis of the gene, other related genes are further introduced to realize that CrtZ and CrtW are positioned in lipid droplets by lipid droplet targeting peptide Olesion, so that the yield of astaxanthin is increased, finally geranyl-geranyl diphosphate synthase, phytoene dehydrogenase and phytoene synthase are targeted to endoplasmic reticulum through endoplasmic reticulum targeting peptide, and the yield of astaxanthin is increased. Therefore, the yield of the fermentation tank is up to 440mg / L. The recombinant saccharomyces cerevisiae provided by the invention is clear in genetic background, can stably and efficiently produce astaxanthin, and has a wide application prospect.

The present application relates to compost fermentation technical field, especially to a kind of compost fermentation device using duck blood protein enzyme.The technical scheme includes: support, jar body and top end top cover rotationally installed on support;Worm, worm wheel are arranged between jar body and support, worm engages worm wheel and one end is provided with hand wheel, can drive jar body rotation angle adjustment.The inner container of jar body is divided into two parts by drain plate;Top cover is provided with motor-driven stirring blade, air inlet pipe and air outlet pipe.The drain valve with elastic plugging element is arranged in the drain outlet of jar body bottom;It is also provided with air pressure control mechanism, including pressure valve on top cover and adjusting valve on air outlet pipe, can be raised by switching adjusting valve state The air pressure in jar, realize air pressure auxiliary drainage.The present application is through angle-adjustable jar body, stirring mixing, air pressure auxiliary drainage and ventilation heat preservation and other structures synergies, make duck blood protein enzyme and material fully contact, maintain stable fermentation environment, realize efficient enzymatic fermentation and percolate convenient discharge, improve compost efficiency and quality, reduce operating intensity.

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 invention discloses a clonorchis sinensis recombinant protease inhibitor Cystatin-2 and a polarization effect of the clonorchis sinensis recombinant protease inhibitor Cystatin-2 on macrophages. The protease inhibitor Cystatin-2 is derived from clonorchis sinensis, can remarkably promote polarization of macrophages, and is an immunomodulatory molecule with application prospects.

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.

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 invention discloses composite enzymolysis oat fermented milk and a preparation method thereof, and belongs to the technical field of dairy product processing. The fermented milk disclosed by the invention is prepared by taking cow milk as a substrate, adding oat enzymolysis powder subjected to compound enzymolysis of protease, cellulase and amylase, and fermenting at 42 DEG C by adopting a compound leavening agent of streptococcus salivarius subsp. Thermophilus, lactobacillus delbrueckii subsp. Bulgaricus, lactobacillus rhamnosus and lactobacillus plantarum. According to the oat fermented milk and the preparation method thereof, oat nutrition is fully released through a composite enzymolysis technology, product characteristics are optimized through synergistic fermentation of the four strains, the oat fermented milk has comprehensive nutrition, smooth taste and functional diversity, and the problems that existing oat fermented milk is insufficient in enzymolysis, poor in strain synergism and insufficient in product stability are solved.

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 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 relates to the technical field of plant growth regulators, in particular to a rooting factor synergistic super-immune protein plant growth regulator, the total weight is 100 parts by weight, and the rooting factor synergistic super-immune protein plant growth regulator is composed of rooting factors, super-immune protein, trace elements, a synergistic aid and a solvent or a carrier; the rooting factor is a compound of indolebutyric acid, naphthylacetic acid and zeatin nucleoside; the super-immune protein is prepared from soybean meal raw materials through protease enzymolysis, heat preservation inactivation at the temperature of 85 DEG C, centrifugation, ultrafiltration purification and concentration. The trace element is a compound of boron, zinc and molybdenum. Through scientific compounding of the rooting factor and the super-immune protein, the synergistic interaction among the components is realized, the rooting of crops can be powerfully promoted, the stress resistance and the immunocompetence can be synchronously improved, and the growth and development of the crops are comprehensively guaranteed.

Compositions and methods useful for the inhibition of cancer metastasis are disclosed.

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.