187results about How to "Rapid purification" patented technology

The invention relates to magnetic fluorescent microspheres and a preparation method thereof. The particle size of the magnetic fluorescent microspheres is 5-10 mu m, the fluorescence excitation wavelength range is 400-700nm, and the fluorescence intensity is not reduced within 24h. The prepration method comprises the following steps: adding a swelling agent into monodisperse carboxylated polystyrene microspheres, and adding magnetic nano microparticles into a swelling system; shaking on a decolorization shaker for 12-48h; using mixed solution of cyclohexane and ethanol for cleaning sediment, and sequentially carrying out ultrasonic dispersion and centrifugal separation till supernatant liquid is colorless under an ultraviolet lamp; and saving a final sediment product in 1ml of liquor. Compared with the existing magnetic fluorescent microspheres, the magnetic fluorescent microspheres have the advantages of uniform and controllable diameter, good fluorescence stability, simple preparation process, multiple types of fluorescence codes and the like, and can not only carry out fast separation and purification on reactants by being applied in the biological macromolecular detection, but also simultaneously detect a plurality of target molecules in a sample to be detected in a reaction tube and a hole, thereby being widely applied in the fields of immunoassay, nucleic acid hybridization, genotype analysis and the like.

The invention provides a method for purifying protein of enzyme aggregate based on self-aggregation short-peptide induction. The method comprises the following steps of: fusing and expressing a target protein, a peptide section (preferably intein) containing a cleavage site and a short peptide with self-aggregation function into a triplet in a sequence from left to right or from right to left, forming an enzyme aggregate in an expressing process, and separating from most soluble impurities in cell lysate by adopting a centrifuging or filtering method; and then releasing the target protein into solution by inducing cutting at a cleavage site so as to achieve the purpose of purification. The method has the characteristics of low cost and simple flow, can be used for laboratory-scale high-throughput protein purification and is also beneficial to industrial-scale protein production.

The invention discloses an SERS (surface-enhanced Raman spectrum) detection method for narcotics in a urine sample. The method comprises steps as follows: synthesizing a sol ultrafine grained nano-material with SERS active noble metals; dropping the sol ultrafine grained nano-material onto a substrate, and obtaining an SERS base through drying; taking the urine sample, sequentially adding a reagent I, namely, an alkaline solution, and a reagent II, namely, solid salt, to the urine sample, adding a reagent III, namely, an organic solvent, after even mixing, and then performing vibrating extraction on the mixture; taking an organic-layer solution after centrifugation or filtration by the aid of an organic filtration membrane; dropping the organic-layer solution onto the SERS base, and performing SERS detection on the organic-layer solution by the aid of a Raman spectrometer. According to the method, the urine sample of a person is pretreated, and the common narcotics in the urine sample are quickly separated, purified and enriched, so that the detection time is shortened, only 3-5 min is taken for the whole detection, and the detection sensitivity and the detection efficiency can be improved.

The invention provides a purification technology for low-grade bentonite, belonging to the field of processing and utilization of non-metal ores. The purification technology comprises the following steps of preprocessing the raw materials; stirring to prepare pulp; ultrasonically purifying; standing for precipitating; separating out upper-layer slurry; removing impurities through centrifuging; filtering and drying. The purification technology is supported by ultrasonic waves; in the presence of a dispersing agent, the impurities can be well separated from montmorillonite, so that the purification effect is improved; furthermore, the purification technology is simple, is small in equipment investment, can be used for purifying the product with high content of montmorillonite from the low-grade bentonite rapidly, efficiently, simply and conveniently, has the advantages of low production cost and short purification period, and is easy to popularize.

The invention discloses high-temperature resistant and alkali resistant bacillus clausii CGMCC No. 1768, a production method thereof, and a method for producing high-temperature alkali protease and other enzyme preparations with the bacterial strain. The bacillus clausii has excellent tolerance, and can grow under extremely adverse environmental conditions as well as in the environment of NaCl of 0 to 20 percent, PH value from 5 to 14, and 4 DEG C to 85 DEG C. The bacillus clausii can survive in wide scope. By adopting the tolerance of the bacterial strain to extremely adverse environment, to research and develop the metabolites of the bacterial strain is very significant. Industrial production requirement can be met; the quality of enzyme preparations can be improved and stabilized; environmental pollution can be reduced; industry can be promoted to be stabilized and developed quickly by developing high temperature resistant and alkali resistant enzymes with excellent thermostability.

The invention discloses a preparation method of dual functional chitosan microspheres. The preparation method comprises the following steps of 1, mixing an ethanol dispersion solution of superparamagnetic Fe3O4 nanoparticles and a deionized water solution of chitosan, and uniformly dispersing the mixture to obtain a mixed solution, 2, through a soybean oil solution of span-80, receiving the mixed solution obtained by electronic injection to obtain a magnetic chitosan microsphere dispersion liquid, and 3, dropwisely adding a cross-linking agent solution into the magnetic chitosan microsphere dispersion liquid and carrying out a reaction process after dropwise addition at a room temperature for 1.5-2.5h to obtain the dual functional chitosan microspheres. The dual functional chitosan microspheres have uniform and controllable sizes and structured morphology. The preparation method utilizes an autofluorescence substance to replace extraneous introduced fluorescence substances such as organic dye and quantum dots thereby avoiding the toxicity produced by the chitosan microspheres on cells in use and inhibiting cell secondary damage.

The invention relates to an acid response water-soluble near-infrared BODIPY light-sensitive agent and a preparation method thereof. The novel BODIPY light-sensitive agent is soluble in water, has absorption performance in a near-infrared area, and has the advantages of being good in water solubility and high in light absorption in the near-infrared area. Meanwhile, the invention further provides the preparation method of the BODIPY light-sensitive agent. Synthesis is quick, and purification is easy. According to the preparation method, by reasonably designing feeding equivalence ratio, the target product can be obtained only through extraction, washing and other simple processing in triglycol monomethyl ether and p-toluenesulfonic acid sulfonylation; by means of the ethyl alcohol re-crystallization method, high-purity BODIPY is obtained; target molecules are synthesized at a high yield through reasonable temperature control. The method has the advantages of being easy to operate, convenient and safe to use and the like. The prepared light-sensitive agent can be soluble in water, the maximum absorption peak is 655 nm, the maximum emission peak is 710 nm, and the light emitting intensity of the light-sensitive agent is gradually improved along with addition of trifluoroacetic acid.

The invention discloses a preparation method of nano magnetic beads for purifying histidine tagged protein. The method comprises the following steps: taking sodium alginate, FeCl3.6H2O, NaAc.3H2O andPEG as raw materials, taking ethylene glycol and deionized water as solvent, reacting through a solvothermal method to generate monodispersed sodium alginate/Fe3O4 nano magnetic beads, subsequently further carrying out an epoxy chloropropane cross-linking reaction and activation on the nano magnetic beads and the newly added sodium alginate, then introducing an iminodiacetic acid (IDA) chelating agent so as to obtain water-stable nano magnetic beads efficiently chelated with metal nickel or cobalt ions; and efficient, rapid, simple and low-cost separation and purification of the histidine tagged protein are realized. The method disclosed by the invention is simple, rapid and stable to operate and low in cost; the synthesized nano magnetic beads are large in specific surface area and stablein a water phase and not prone to agglomeration and precipitation; the purification efficiency of the histidine tagged protein is greatly improved; and the large-batch industrial production of the nano magnetic beads can be realized.

The invention aims to provide a method for simply and quickly purifying genomic deoxyribonucleic acid (DNA) of plant tissues by using a magnetic nano composite material. The method comprises the following steps of: performing sample lysis by using two kinds of lysis solution, wherein the first lysis solution comprises PVP (poly vinyl pyrrolidone) with mass-to-volume ratio of 2 to 5 percent, 2 to 5 percent of CTAB (cetyl trimethyl ammonium bromide), and beta-mercaptoethanol with volume fraction of 0.5 to 2 percent, and the second lysis solution comprises 3 to 6M of guanidine hydrochloride and Triton X-100 with volume fraction of 2 to 5 percent; blending, and performing full lysis for 5 to 30 min in a water bath; and adding 10 to 50 microliters of 10 mg/ml RNase solution, and thus obtainingthe sample lysis solution with the genomic DNA. The purification method is easy to operate, has quick purification process, and has the advantages of high purification rate, high DNA integrity, high purity and the like. According to the method, fractional tissues lysis is not required, and the requirement of extracting the genomic DNA of various plants and different places is met to the great extent.

The invention belongs to the field of applied molecular biology, and particularly relates to a virus-like particle containing novel coronavirus nucleic acid as well as a preparation method and application of the virus-like particle. The virus-like particle containing the novel coronavirus nucleic acid contains MS2 bacteriophage coat protein and a recombinant gene S sequence wrapped in the coat protein, wherein the S sequence consists of a novel coronavirus ORF1ab gene segment, a human beta-Globin gene segment, a novel coronavirus N gene segment and an E gene segment which are connected in sequence; and the S sequence has a nucleotide sequence as shown in SEQ ID NO.5. The virus-like particle can be used for whole-course quality control of nucleic acid extraction, amplification and detectionof the novel coronavirus, and can be popularized and applied as a positive control and internal standard quality control product of the novel coronavirus in a kit.

The invention relates to a recombinant beta-glucosidase and an expression and purification method and the immobilization application thereof, and belongs to the technical field of biological enzyme engineering. The recombinant beta-glucosidase is recombinant beta-glucosidase Glu-linker-ELP-GB (GLEGB) obtained by linking binary labels ELP and GB with beta-glucosidase (Glu) through a universal linking peptide linker, wherein the sequence of GB polypeptide is linked to the 3' end of an ELP gene. By virtue of a reversible phase change property of an ELP label on recombinant enzyme, separation andpurification of the enzyme can be achieved; the recombinant enzyme containing the ELP label can be precipitated only by adding a certain amount of (HN4)2SO4 for incubation for a period of time; by virtue of a temperature response behavior of the ELP label, one-step rapid separation and purification of a target enzyme molecule is achieved; the enzyme purifying effect of the expression and purification method is superior to that of an ammonium sulfate precipitation method, and the purification cost of the expression and purification method is much lower than that of a chromatography method.

The invention belongs to the field of nucleic acid purification, and in particular relates to a method for purifying total nucleic acid (comprising genome DNA and total RNA) by using gold-magnetic particles. The method comprises the following steps: preparing sample lysing solution containing the total nucleic acid; mixing the gold-magnetic particles and the biological sample lysing solution to form a gold-magnetic particle-total nucleic acid compound under the action of binding buffer solution; magnetically separating and cleaning the gold-magnetic particle-total nucleic acid compound; and finally, eluting the total nucleic acid from the surfaces of the gold-magnetic particles by using eluent, and collecting the supernate after magnetic separation to obtain purified total nucleic acid solution. According to the purifying method, centrifugation is saved, so the operation is simple and convenient, the purifying process is quick, the quality of the obtained total nucleic acid is high and the cost is low, and the genome DNA and the RNA in the biological sample can be obtained by separating at the same time.

The invention discloses a foreign protein soluble expression plasmid, a preparation method thereof and application method thereof, which belong to the technical field of bioengineering. The plasmid is pET30E, wherein the DNA sequence of the plasmid is presented by Seq ID No.1. An amplified product obtained by performing PCR amplification on a plasmid template and a primer group is connected with a pMD18-t vector; an Escherichia coli clone is obtained by transforming Escherichia coli DH5 alpha; and a product obtained by performing enzyme cutting connection on the Escherichia coli clone serving as a substrate is used for transforming the Escherichia coli DH5 alpha to form soluble expression plasmid molecules. The application method comprises the following steps that: the obtained expression plasmid molecules are used to transform Escherichia coli BL21 (DE3) competent cells through thermal activation to form a recombinant Escherichia coli colony; and after a bacteroid cell is obtained through inoculation culture, the soluble expression of the expression plasmid molecule is improved. The plasmid of the invention has promotion effect on assistance of the soluble expression of foreign proteins in Escherichia coli and has a great significance for further analysis of the functions of the proteins.

The present invention relates to a production method of tea leaf; the tip leaf of weeping forsythia capsule which is a green forsythia plant is taken as the raw material; the method comprises the following steps that the tip leaf of weeping forsythia capsule is picked and cleaned; the leaf is spread for 5 to 12 hours; the leaf is fried and steamed and the water is removed with a temperature of 120 to 180 Celsius degrees; the leaf is kneaded primarily for 8 to 12 minute with a temperature of 40 to 50 Celsius degrees; the leaf is arranged or formed with a temperature of 65 Celsius degrees; the leaf is baked with a temperature of 70 to 80 Celsius degrees; the leaf is kneaded again for 5 to 6 minutes with a temperature of 40 to 50 Celsius degrees; the leaf is baked again with a temperature of 60 to 80 Celsius degrees and the temperature is changing from a higher one to a lower one; the leaf is baked until the peduncle is broken when the peduncle is removed and the leaf becomes powder when the leaf is kneaded with hand. The beneficial effect of the present invention is that the fresh lead is heated evenly without local high temperature which can be filtrated through the flesh leaf of leaf pile in a short time; the oxydase is purified fast and the chlorophyll is preserved to the most; therefore, the tea is greener and fresher; the taste is diluted and the color is strong; the tea can be stewing for a long time.

The invention belongs to the technical field of tea processing technology and particularly relates to a tea processing method including following steps: picking, cleaning, tedding, de-enzyme, cooling, rolling, drying, selecting and packaging. By means of the tea processing method, fresh tea is heated uniformly without local high temperature. Foliages or stacks of fresh leaves can be penetrated in a short time. Oxidase can be quickly passivated so that chlorophyll can be retained maximumly. The tea is more green and fresh, is delicate in taste, is thick in color and is soaking-durable.

A process for preparing the hybrid seeds of rape and the vegetables in the mustard family by using the sterile cytoplasm of charlock includes such steps as separating the protoplast from leaf pulp, purifying it, pre-treating, fusing, culturing, discriminating the hybrid, reproduction, selectively culturing the male sterility recovery line of charlock cytoplasm, transferring the male sterility and recovery characters from the charlock cytoplasm to rape or the vegetables in mustard family, and hybridizing between the charlock cytoplasm male sterility line of rape or the said vegetables and the male fetile rape or the said vegetables.

The invention provides a method for preparing recombinant human interleukin-11 (rhIL). The method comprises the following steps: 1) providing a fusion protein, wherein from the N-end to the C-end, the fusion protein is thioredoxin-(His)6-proteolytic enzyme recognition site-rhIL-11 or (His)6-thioredoxin-proteolytic enzyme recognition site-rhIL-11; 2) using the proteolytic enzyme to perform enzyme cutting to the fusion protein and obtain an enzyme cutting product, wherein the enzyme cutting product contains thioredoxin and rhIL-11; and 3) using the nickel ion chelate affinity column chromatography to purify the enzyme cutting product, and performing gradient elution to the column to obtain rhIL-11, wherein the thioredoxin and the rhIL-11 are both adsorbed on the column. By adopting the method provided by the invention, the rhIL-11 can be purified rapidly, conveniently and efficiently and the recovery rate can be greatly increased.

The invention discloses a method for expressing and purifying recombinant ethanol oxidase rAOX. According to the method, an ethanol oxidase-poly-L-histidine expression vector pPIC9K-AOX-HIS is constructed first by a molecular biological technique, then the expression vector pPIC9K-AOX-HIS is linearized and transferred to pichia pastoris GS115, and thus, pichia pastoris engineering bacteria capable of expressing active recombinant ethanol oxidase are obtained, and part of recombinant ethanol oxidase can be secreted and expressed. Through optimization of the expression conditions of recombinant ethanol oxidase, the ethanol oxidase produced by the engineering bacteria may reach 1,862U/L. Meanwhile, the secretion and expression of the ethanol oxidase and the introduction of a poly-L-histidine tag simplify the purification process of the recombinant ethanol oxidase. According to the result of tests, the recombinant ethanol oxidase expressed by the engineering bacteria has much higher thermostability than pichia pastoris wide type ethanol oxidase. The operation in the whole process of the method is simple, so the method is suitable for industrial production and can create certain economic benefit.

The invention provides a rapid purification reagent for sputum microorganisms. The rapid purification reagent is prepared from a liquefaction solution, cell lysate and an exonuclease buffer solution,wherein the cell lysate contains guanidine salt and a surface active agent, and the concentration of the guanidine salt in the cell lysate is 1-3mol/L. The invention also provides application of the rapid purification reagent for the sputum microorganisms. The rapid purification reagent, provided by the invention, for the sputum microorganisms is prepared by combining low-concentration guanidine hydrochloride with the surface active agent and a reducing agent for completely lysing human cells (nucleus), and genomic DNA is released into a solution due to cell disintegration, most of the DNA isremoved during centrifugal obtaining of bacteria, and the residual free DNA is further digested by DNase; the bacteria are not affected or seldom affected due to the protection of the cell wall.