531 results about "Agarose" patented technology

The invention relates to the technical field of crop molecular genetic markers, and discloses an InDel molecular marker of avena nuda and application thereof.The molecular marker is a group of 314bp deletion variation and 2bp insertion variation in the 610439484-610440026 interval of the 5C chromosome position of a reference genome of an avena nuda variety OT3098, the nucleotide sequence of the deletion variation is shown as SEQ ID NO.1, the nucleotide sequence of the insertion variation is shown as SEQ ID NO.2, and the nucleotide sequence of the insertion variation is shown as SEQ ID NO.3. The nucleotide sequence of the insertion variation is 5 '-AA-3'. The molecular marker provided by the invention can realize stable detection in agarose gel electrophoresis by combining a specific primer pair (SEQ ID NO.2-3) with an optimized touchdown PCR (Polymerase Chain Reaction) program, has the advantage of being not influenced by a tissue part and a growth period, and provides an efficient and reliable technical means for oat germplasm identification, seed purity detection and molecular breeding.

The invention discloses a fruit and vegetable exosome extraction method based on ligand fishing. The fruit and vegetable exosome extraction method comprises the following steps: suspending amino-functionalized Fe3O4 (at) SiO2 nanoparticles in a glutaraldehyde solution, and adding activated agglutinin to obtain Fe3O4 (at) SiO2-agglutinin nanoparticles; adding a glutaraldehyde solution into agarose gel to obtain a size exclusion chromatographic column filler; the method comprises the following steps: dispersing Fe3O4 (at) SiO2-lectin nanoparticles in a PBS buffer solution to obtain a uniform suspension, adding the uniform suspension into a size exclusion chromatographic column filler to obtain a uniform slurry, filling the size exclusion chromatographic column with the uniform slurry, and flushing with the buffer solution to form a magnetic bead stationary phase; and loading fruit and vegetable supernatant into a size exclusion chromatographic column, passing through the magnetic bead stationary phase, cleaning residual impurities combined on the exosome-magnetic bead compound by using a buffer solution, and dissociating the exosome from the magnetic beads by using a competitive eluent of the exosome to obtain the high-purity fruit and vegetable exosome. The problem that an existing exosome extraction method is difficult to meet the requirements of high efficiency, simplicity, high purity and high activity at the same time is solved.

The invention discloses a fluorescence ratio type carbon dot hydrogel sensor, a preparation method thereof and application of the fluorescence ratio type carbon dot hydrogel sensor in rapid detection of doxorubicin hydrochloride. The invention discloses a fluorescence ratio type carbon dot hydrogel sensor which is characterized in that 3-aminophenol is used as a carbon source, absolute ethyl alcohol is used as a solvent, carbon dots are prepared through a one-step solvent method, the carbon dots have green fluorescence, agarose is dissolved in a mixed solution of deionized water and a phosphate buffer solution to be fully dissolved, and then the carbon dots are prepared into the fluorescence ratio type carbon dot hydrogel sensor. And adding the carbon dots into the mixed solution, fully dispersing, and cooling to obtain the fluorescence ratio type carbon dot hydrogel sensor. The sensor disclosed by the invention has the characteristics of high selectivity and high sensitivity when being used for detecting doxorubicin hydrochloride, and also has the characteristics of convenience, visibility, rapidness, portability, easiness in operation and the like.

The invention discloses bioactive macroporous hydrogel as well as a preparation method and application thereof. The bioactive macroporous hydrogel comprises hydrogel particles and a bioactive load loaded on the hydrogel particles, the hydrogel particles are obtained by mechanically extruding a hydrogel matrix, and the hydrogel matrix is formed by compounding sodium alginate, agarose and methacrylated hyaluronic acid through hydrogen bond crosslinking and photo-crosslinking; the bioactive load comprises: a) small extracellular vesicles of which the surfaces are modified with macrophage targeted CRV peptides; and b) a PLGA (poly (lactic-co-glycolic acid)) microsphere loaded with a cartilage inducer Kartogenin. The bioactive macroporous hydrogel disclosed by the invention has an internally communicated macroporous network, is beneficial to cell infiltration, blood vessel ingrowth and nutrient substance diffusion, and overcomes the defects that the traditional hydrogel is small in pore size and limits cell behaviors. According to the bioactive macroporous hydrogel disclosed by the invention, dual bioactive loads are adopted to synergistically promote tendon-bone healing, so that the treatment effect is better.

The invention belongs to the technical field of plant fungus molecular biology detection, and discloses an LAMP primer group, a kit and a detection method for detecting oat smut pathogenic bacteria. A group of LAMP (loop-mediated isothermal amplification) specific primers are designed according to a specific sequence on a whole genome of the oat smut pathogenic bacteria Ustilago hordei, results are judged through a real-time fluorescence quantification method, an agarose gel electrophoresis method and an SYBR Green I fluorescent dye developing method, and the oat smut pathogenic bacteria carried by oat seeds are detected. The LAMP detection method for the oat smut pathogenic bacteria, established by the invention, is strong in specificity, high in sensitivity, high in speed and low in cost, provides a new technical means for detection of the oat smut pathogenic bacteria carried by the oat seeds, and has relatively high practical application value.

The invention discloses a device and method for automatically shearing and recycling agarose gel, and relates to the technical field of agarose gel cutting, and the device and method for automatically shearing and recycling agarose gel comprise an image acquisition unit, a three-axis moving mechanism, a gel cutting device, an electrophoresis apparatus and a control unit; the image acquisition unit comprises a light source assembly and an image acquisition camera corresponding to an irradiation area, the image acquisition camera is used for shooting a dyed electrophoresis image, and the light source assembly is used for obliquely irradiating gel to enable a sample strip to develop color; the control unit comprises an analysis processing unit; the analysis processing unit comprises a deep learning detection module and a data processing module; the control unit cooperates with the image acquisition unit, the three-axis moving mechanism and the gel cutting device, so that a complete automatic process from strip identification and positioning after electrophoresis to gel block cutting, grabbing, releasing and recycling is constructed.

The present application discloses a specific DNA fragment for sex identification of Pelteobagrus ussuriensis and a method for sex identification, specifically relating to the technical field of sex identification of Pelteobagrus ussuriensis. The above-mentioned identification method includes the following steps: Dissect 5 female and 5 male Pelteobagrus ussuriensis of a full-sib family to determine the physiological sex; respectively extract DNA from the muscles of female samples and male samples, construct a paired-end genomic DNA library with an insert fragment size of 300 bp, and perform high-throughput sequencing; based on the GWAS and Fst methods, identify that the sex chromosome of Pelteobagrus ussuriensis is chromosome 8, and then screen for Indels within the sex determination region on chromosome 8; screen a 601-bp male-specific DNA sequence and design and synthesize primers; use PCR amplification and agarose gel electrophoresis detection, and the result without specific bands is female, and the one with a 356-bp band is male. This scheme has the advantages of strong specificity, simple primer composition, and more intuitive identification of the sex of Pelteobagrus ussuriensis.

The invention provides a recombinant III-type humanized collagen gel based on a macromolecular crowding effect as well as a preparation method and application of the recombinant III-type humanized collagen gel. The method comprises the following steps: firstly, preparing an rhCol III solution system; then preparing a macromolecular crowding bath containing dextran 70, polyethylene glycol 8000 and ascorbic acid; then adding an rhCol III solution into the macromolecular crowding bath to form an rhCol III microgel; and finally, constructing a 3D printing molecular exclusion mold system which takes agarose as a substrate and contains a functional additive and a temperature-sensitive polymer as a temperature-sensitive release and spatial constraint component, so as to realize directional construction of the rhCol III microgel. According to the method disclosed by the invention, second-level physical gelling (lt; 15s) of rhCol is realized, and a physiological microenvironment supporting cell proliferation is constructed. The recombinant III-type humanized collagen gel prepared by the method can be used for preparing collagen microgel balls, collagen bundles, 3D (three-dimensional) bio-printing products and the like, and is wide in application range.

The invention belongs to the technical field of cosmetics, and particularly relates to a water-in-oil-in-water three-phase coated microbead as well as a preparation method and application thereof. The water-in-oil-in-water three-phase coated microbead is of a spherical structure and sequentially comprises an outer water phase, an inner oil phase and an inner water phase from outside to inside, the outer water phase comprises sodium alginate, agarose, cyclodextrin, glycerin and other components, the inner oil phase comprises caprylic / capric triglyceride, grape seed oil, olive squalane, tocopherol and other components, and the inner water phase comprises sodium alginate, agarose, cyclodextrin, glycerin and other components. And the inner water phase comprises ampelopsis grossedentata extract, astragalus membranacus root extract and other components. According to the invention, an easily oxidized and unstable active matter is fully and firmly protected by adopting a droplet microfluidic coating technology, so that the active matter is free from external stimulation, active ingredients with different effects play multiple roles, and the composition has good anti-inflammatory, soothing and oil-controlling effects and can be applied to the field of cosmetics.

The invention discloses light-driven toxin-enriched composite hydrogel as well as a preparation method and application thereof. The composite hydrogel comprises light-driven hydrogel and detection hydrogel, the light-driven hydrogel is agarose hydrogel doped with Au (at) Ag core-shell nano particles; the detection hydrogel is a double strand formed by an okadaic acid aptamer and a complementary sequence cDNA thereof, and a double strand formed by a domoic acid aptamer and a complementary sequence DNAzyme thereof, the hairpin H1 is used for modifying a quenching group BHQ2 at the 3'end, the hairpin H2 is used for modifying a fluorophore Cy3 at the neck part, the hairpin H3 is used for modifying a fluorophore FAM and a quenching group BHQ1 at the two ends respectively, and the metal ion doped agarose hydrogel is used for catalyzing DNAzyme cyclic shearing; and the photo-thermal driving unit is positioned on the detection function unit. According to the present invention, with the composite system of upper layer photo-thermal enrichment and lower layer dual-signal detection, the rapid and high-sensitivity simultaneous detection of the okadaic acid and the domoic acid is achieved through the combination of the photo-thermal transpiration effect and the HCR and DNAzyme cyclic shearing reaction;

The invention is applicable to the technical field of polymer biomedical hydrogel materials, and provides a preparation method and application of multifunctional agarose-based particle composite hydrogel based on liquid-liquid phase separation, and the agarose-based particle composite hydrogel is a product formed by introducing microsphere particles into an agarose hydrogel body matrix; the microsphere particles are formed by a prepolymer and sodium lignin sulfonate through phase separation mediated by a composite behavior based on non-covalent interaction; the prepolymer is obtained by carrying out amine-epoxy reaction on an amino compound and an epoxy compound in water. According to the prepared agarose-based particle composite hydrogel, a microsphere particle structure is formed on the surface of an agarose matrix while positively charged amino groups and sodium lignin sulfonate are introduced, so that the multifunctional agarose-based particle composite hydrogel is prepared.

The invention provides a high-flux porous microsphere for chromatography and a preparation method thereof, and the preparation method of the high-flux porous microsphere for chromatography comprises the following steps: respectively dissolving agarose and aminated glucan in water, and uniformly mixing an agarose solution and an aminated glucan aqueous solution to obtain an aqueous phase; adding the water phase into the oil phase containing the emulsifier, and stirring and emulsifying at 70-90 DEG C for 30-90 minutes to obtain a mixed solution; cooling the obtained mixed solution to 20 DEG C or below at a speed of 1-5 DEG C / min, and curing into a ball to obtain a microsphere; and adding NHS-PEG-COOH into the obtained microspheres, carrying out a primary cross-linking reaction, adding epoxy chloropropane into the obtained microspheres, and carrying out a secondary cross-linking reaction to obtain the high-flux porous microspheres for chromatography. According to the preparation method of the high-flux porous microspheres for chromatography, the separation efficiency of the obtained porous microspheres can be improved, and relatively high dynamic adsorption capacity can still be realized at a high flow rate.

The invention provides an adsorption material and a preparation method and application thereof, the adsorption material comprises a carrier and an endotoxin affinity binding ligand, the endotoxin affinity binding ligand comprises a recombinant C factor protein, and the carrier and the recombinant C factor protein are connected through a spacer arm; wherein the carrier comprises at least one of resin, chitosan, cellulose, agarose and glucan; the spacer arm comprises at least one of modified polyethylene glycol, polyvinyl pyrrolidone, polymethylacrylic acid-2-hydroxyethyl ester, polyvinyl alcohol, polyacrylic acid, hexamethylenediamine, divinyl sulfone and glutaraldehyde, and at least one end of the modified polyethylene glycol is provided with carboxyl, aldehyde or amino. The adsorption material provided by the invention can efficiently adsorb endotoxin, has the characteristics of low toxicity risk and excellent blood compatibility, and can improve the safety of blood purification.

The invention discloses KASP primers for identifying genotypes of chicken green-shell eggs and application of the KASP primers. Compared with a common flow of identifying genotypes through PCR (Polymerase Chain Reaction) and agarose gel electrophoresis, the method for performing KASP genetic typing by utilizing the primers with the sequences as shown in SEQ ID NO: 1-3 provided by the invention has the advantages that the steps of preparing agarose gel and performing electrophoresis are omitted, and the economic cost is reduced; when large-scale detection is carried out, a large amount of manpower resources are saved, and the detection time is shortened; analysis is directly carried out according to an instrument detection result, personal errors are reduced, and the accuracy of a typing result is improved.

The invention discloses a degradable preservative film and a preparation method thereof. The film takes nano chitosan / agarose in a ratio of 1: 1 as a matrix, contains 0.5-5wt% of PVA (Polyvinyl Alcohol), 1-4wt% of protein powder or starch, 0.2-3wt% of tea polyphenol / carvacrol, 5-20wt% of glycerol and optional 5-15wt% of polylactic acid, and is cross-linked with sodium alginate. And carrying out tape casting at 55-65 DEG C, and carrying out gradient drying at 40-50 DEG C, thereby obtaining the product. The film is compact and flexible, the breaking strength is larger than or equal to 69 MPa, the oxygen permeability is low, the inhibition zone on six germs is enlarged by 3-4 mm, 98% of natural soil is degraded 20 days later, and no toxic smoke is generated during incineration. The biodegradable film is solvent-free in preparation, can be produced by existing equipment, is lower than a traditional biodegradable film in cost, can be in direct contact with food, remarkably prolongs the fresh-keeping period of fruits and vegetables, replaces PE and PVC, and solves white pollution.

The invention discloses a toning lotion composition, and belongs to the technical field of cosmetics. The toning lotion composition is prepared from the following preparation raw materials in percentage by mass; 0.09%-0.23% of a thickening agent, 2%-9% of an oily component, 0.1%-2% of a stabilizing agent and the balance of water; the thickening agent is prepared from agarose, gellan gum and xanthan gum; the stabilizing agent is selected from at least one of sodium chloride, calcium chloride or disodium EDTA (Ethylene Diamine Tetraacetic Acid); the viscosity of the toning lotion composition at the temperature of 25 DEG C is 50 to 500 Pa.s. The toning lotion has fresh and cool use feeling and moist degree of oil, is high in transparency and flexible in use mode, can be used as essence for smearing and can also be atomized through a spray bottle.

The invention relates to porous ceramic for a high-air-permeability semiconductor device as well as a preparation method and application of the porous ceramic, raw materials comprise 35-55 wt% of alumina powder and 10-55 wt% of a polymer microsphere pore-forming agent, and the polymer microsphere pore-forming agent comprises one or more of agarose, polylactic acid, polyacrylamide and organic silicon resin. The porous ceramic for the high-air-permeability semiconductor device, which is obtained by the preparation method provided by the invention, has relatively high porosity and air permeability, and also has relatively high mechanical strength and dielectric strength. According to the present invention, the gas volume flow rate is applied to the internal pore size characterization, and the high porosity and the high gas permeability can be achieved by accurately measuring the gas flow transmittance, accurately characterizing the pore size and the opening degree in the porous ceramic, and adjusting the particle size and the addition amount of the polymer microsphere pore forming agent; through the preparation process, the porosity and mechanical strength of the porous ceramic for the high-air-permeability semiconductor device can be balanced, meanwhile, high air permeability and high compressive strength are obtained, and the porous ceramic is particularly suitable for electrostatic chucks of different manufacture procedures.

The invention discloses an InDel molecular marker related to seedless traits of filial generations of grapes and application of the InDel molecular marker. The nucleotide sequence deletion part of the InDel molecular marker for the seedless type of the filial generation of seedless grape and nucleated grape is shown as SEQ ID NO.3, and is positioned at the upstream 263-693 bp of a VviAGL11 gene transcription start site (TSS) which is closely linked with the seedless character of the grape, and the nucleotide sequence deletion part of the InDel molecular marker is shown as SEQ ID NO.6. The nucleotide sequence deletion part of the InDel molecular marker is shown as SEQ ID NO.7. The nucleotide sequence deletion part is shown as SEQ ID NO.8. According to the invention, PCR (Polymerase Chain Reaction) amplification and Sanger sequencing are carried out on VviAGL11 gene promoters of a seedless grape variety 'Zhongqing 18' and a seedless grape variety 'Musang', so that polymorphic sites with significant difference are obtained, and the difference between the polymorphic sites is 431 bp. The InDel molecular marker is suitable for grape seedless molecular marker-assisted selective breeding, and the invention also discloses a method for carrying out genotype identification through an agarose electrophoresis technology in a seedling stage of hybrid generations of seedless grapes and seedless grapes, so that early selection of seedless characters of the grapes is realized; the seedless grape breeding efficiency is further improved, the breeding cost is reduced, and the breeding process is promoted.

The invention discloses a method for cultivating triploid kelp sporophytes and identifying ploidy of induced protonema, and belongs to the field of kelp breeding. The method is based on a capillary glass needle separation technology for inducing filamentous somatic cells by kelp and a molecular biological technology for specific marking of sex of kelp gametophytes. The sex specific marker of the kelp gametophyte is used for carrying out PCR (Polymerase Chain Reaction) amplification and 1.5% agarose gel electrophoresis detection on the separated induction protonema single-cell cloning line, so that the genetic sex of the kelp induction protonema can be accurately identified; and the ploidy of the induced protonema of a part of varieties (lines) is directly determined from the molecular level. The method for synchronously detecting the genetic sex and ploidy of the induced protonema breaks through the bottlenecks of chromosome counting and flow cytometry operation, can be completed only through one-time PCR amplification, and is simple, convenient, rapid, short in time consumption, capable of completing detection of a plurality of samples at the same time and low in cost.

The invention relates to the technical field of microsphere modification, in particular to an agarose magnetic bead and a preparation method thereof, a functionalized agarose magnetic bead and a preparation method thereof, and the preparation method of the agarose magnetic bead comprises the following steps: dissolving agarose in water to obtain an agarose solution; and heating the agarose solution to boiling, adding the magnetic bead solution and the emulsion, stirring, cooling with cold water, and continuously stirring to obtain the agarose magnetic beads. According to the preparation method, the agarose solution and the magnetic bead solution are stirred together, so that the magnetic beads and the agarose are wrapped together, the agarose magnetic beads with the particle size ranging from 10 micrometers to 200 micrometers are synthesized under the action of the emulsifier, and the agarose magnetic beads prepared through the preparation method have the advantages of being high in magnetism, uniform in particle size distribution, simple in equipment and easy to produce on a large scale. Moreover, a suspension stirring method is utilized, the magnetic beads are coated in the agarose, so that the agarose has magnetism, and the agarose magnetic beads with relatively good uniformity can be prepared.

The present application belongs to the field of molecular genetics, and particularly relates to an ethylene response transcription factor HvERF62 for regulating the moisture tolerance of barley and application thereof in molecular marker assisted selection breeding. A site significantly affecting the moisture tolerance of barley is identified through whole genome association analysis, and a candidate gene is predicted, and further, the moisture tolerance function of the candidate gene HvERF62 is verified by using the CRISPR-Cas9 gene editing method. The function of HvERF62 for regulating the moisture tolerance of barley is determined by using the gene editing technology, and an InDel molecular marker for detecting the moisture tolerance of barley is developed. The molecular marker of the present application is detected by using 3% agarose gel, is stable, simple in method, and can be used for screening the moisture tolerance of barley at an early stage, and has a strong application value in the breeding of the moisture tolerance of barley.

The invention discloses a preparation method of a novel composite carbon source. The preparation method comprises the following steps: S1, preparing a solution; s2, preparing a culture medium; s3, extracting a genome; s4, extraction of plasmid DNA (deoxyribonucleic acid); s5, preparation of agarose gel electrophoresis; s6, constructing a linearized vector plasmid; s7, preparing an electrotransfection competent cell; and S8, electric conversion treatment. According to the preparation method of the novel composite carbon source, pfkA and pfkB can be respectively, independently and simultaneously knocked out when the carbon source is prepared by utilizing a gene editing technology, the consumption of a glucosamine precursor 6-fructose phosphate in an EMP pathway can be reduced, more glucose is used for synthesizing a target product, and by optimizing the preparation process of the composite carbon source, the yield of the target product is improved. The shake-flask fermentation yield of the glucosamine can be obviously improved, the conversion rate of the composite carbon source is also improved, and cell growth and synthesis of the target product glucosamine can be reasonably balanced, so that the yield of the glucosamine is improved.

The utility model belongs to the technical field of sepharose gel preparation, and particularly relates to a temperature measuring and blending device for sepharose gel preparation, which comprises an oscillating and blending device, the oscillating and blending device comprises a vibrating plate capable of vibrating, a fixing device for fixing a preparation container is clamped on the vibrating plate, and a temperature measuring device is arranged on the vibrating plate. The fixing device is fixedly connected with a temperature measuring plate for placing a preparation container, and a thermochromic film is arranged on the upper surface of the temperature measuring plate. Compared with the prior art, by arranging the fixing device and the temperature measuring plate on the fixing device, the technical problems that in the prior art, an agarose solution needs to be shaken up manually, and the agarose solution is possibly polluted when a traditional temperature measuring device is used are solved.

The application provides a kind of construction and application for sesbania sesban DNA fingerprint, belong to sesbania molecular marker technical field, the application is based on sesbania genome sequencing, first develops and screens out 8 pairs of SSR primers with sequence specificity and rich polymorphism, by analyzing the agarose gel electrophoresis result of PCR amplification product of SSR primer in different varieties, the assignment of different size, type of PCR amplification product is carried out, to form the DNA fingerprint of each sesbania, the application has the characteristics of easy operation, good repeatability, simple data processing, low cost and the like, and lays a foundation for the classification, identification and utilization of sesbania germplasm resources.

The application belongs to the field of fish development, and particularly relates to a molecular marker for identifying Nibea albiflora sperm-induced Pseudosciaena heteroclita gynogenesis offspring and hybrid offspring, a primer set thereof, a kit and application. The primer set for identifying the molecular marker of the Nibea albiflora sperm-induced Pseudosciaena heteroclita gynogenesis offspring and hybrid offspring comprises primers F and R, the nucleotide sequence of the primer F is shown as SEQ ID NO. 1, and the nucleotide sequence of the primer R is shown as SEQ ID NO. 2. Based on genomic data analysis, the application develops the molecular marker F / R which can effectively identify the Pseudosciaena heteroclita gynogenesis offspring and hybrid offspring in the juvenile stage. The method comprises extracting genomic DNA of parents and offspring, PCR amplification and agarose gel electrophoresis detection, and finally distinguishing the gynogenesis individuals and hybrid individuals by whether the Nibea albiflora parent marker is contained in the electrophoresis band.

A primer pair, method, and use for rapidly identifying newborn nude mice. On the basis that a deletion mutation occurs in the base G at position 337 of a coding region of the Foxn1 gene in nude mice, a primer pair introducing an enzyme digestion site is designed according to sequence characteristics. Using the genome of a mouse to be tested as a template, the Foxn1 gene is amplified according to a PCR-RFLP method. The PCR product is then digested with a SmaI restriction endonuclease, and the digested product is genotyped by agarose gel electrophoresis. In addition, a reaction system that can be combined with lateral flow technology is screened, thereby achieving instant detection of the newborn nude mice. The method has the characteristics of simple operation, short time consumption, low cost, instant detection, etc.

This invention discloses the application of green tea acidic polysaccharide GTPS in improving metabolic syndrome, aiming to provide a new use for green tea acidic polysaccharide GTPS. The extraction process involves first grinding green tea into powder, boiling, filtering, deproteinizing using the Seval method, concentrating, adding ethanol, allowing it to stand overnight, centrifuging to collect the precipitate, dissolving it, removing the ethanol, and freeze-drying to obtain GTPSs. The GTPSs are then passed through a DEAE-agarose FF column, eluted with distilled water and 0.2, 0.5, and 1.0 mol / L sodium chloride, and the fractions are collected. The fraction eluted with 0.2 mol / L sodium chloride is dialyzed to desalt, yielding green tea polysaccharide GTPS. Applying this to the improvement of metabolic syndrome provides a solid foundation for revealing the key active components in green tea that promote metabolism and paves the way for the future utilization of tea polysaccharides in the pharmaceutical and food industries, offering a promising solution for the integration of tea resources and the application of natural products.

The invention belongs to the technical field of bioengineering, and particularly relates to a method for purifying single-stranded DNA and application thereof. The invention provides a method for purifying single-stranded DNA (deoxyribonucleic acid). The method comprises the following steps: S1, carrying out polymerization reaction on a primer of which the 5'end is modified with an acrylamide group to generate a linear polyacrylamide modified primer LPA-primer; s2, by taking the LPA-primer as a primer, carrying out PCR (Polymerase Chain Reaction) amplification, so as to obtain a double-stranded DNA (Deoxyribose Nucleic Acid) NALPA-dsDNA (Deoxyribose Nucleic Acid) crosslinked by polyacrylamide; s3, carrying out denatured agarose gel electrophoresis on the LPA-dsDNA under an alkaline condition, and separating a target ssDNA from a non-target ssDNA by utilizing a mobility difference caused by a polyacrylamide modified group; and S4, carrying out gel cutting and recycling on the target ssDNA strip to obtain the purified ssDNA. The result of the embodiment shows that the method can effectively improve the purity of the single-stranded DNA and reduce the cost.

This invention provides a method and kit for enhancing the yield and quality of gene synthesis products, comprising the following steps: providing a total primer mixture, which includes multiple oligonucleotide single strands encoding a target gene sequence / encoding an operational sequence and sterile water; using the primer mixture as a template, performing the first... PCR The reaction yielded the primary spliced ​​product; a second reaction was then performed using start and end primers. PCR The reaction yielded the crude gene synthesis product; nested primers were then used for a third reaction. PCR The reaction yields a nested structure. PCR Products; nested PCR The product was subjected to agarose gel electrophoresis, and the target band was excised and recovered. This application improves upon traditional gene synthesis methods by adding operation sequences to both ends of the synthesized gene sequence and utilizing highly specific nested primers for nested gene synthesis. PCR It can synthesize more than 2000bp in one go. DNA Double-stranded sequences significantly improve the yield and quality of gene synthesis products.

The invention discloses an efficient purification process of a bispecific antibody and a special chromatography system thereof. The efficient purification process of the bispecific antibody comprises the following steps: pretreatment: performing 0.22 mu m microfiltration on a cell culture supernatant to remove cell debris, and performing deep filtration to reduce turbidity to be less than or equal to 1NTU so as to reduce subsequent chromatography medium pollution; carrying out improved Protein A affinity chromatography, namely carrying out affinity chromatography by adopting an agarose medium modified by a recombinant Protein A ligand, taking 20mM Tris-HCl as a loading buffer solution, and taking the pH value of the loading buffer solution as 7.4, taking 100mM citric acid as an elution buffer solution, and taking the pH value of the elution buffer solution as 3.2-3.5; performing mixed mode chromatography: adopting a composite mode medium containing a benzamidine functional group, taking 50mM sodium phosphate as a buffer solution, adjusting the pH value to 6.0-6.5, adjusting the NaCl concentration to 0.1-0.3 M, and specifically removing a double-antibody polymer and a mismatched isomer by utilizing a hydrophobic effect and an ion exchange synergistic effect; and performing nanoscale filtration and terminal treatment: removing viruses by adopting a virus filtration membrane with the aperture of 30nm, and performing 0.22 mu m sterile filtration to obtain a final product. According to the process, the purification efficiency, the yield and the product purity are remarkably improved, and the process is suitable for efficient purification of the bispecific antibody.