116results about How to "Large amount of sample" patented technology

Methods for highly parallel Sanger sequencing are discussed. In particular, provided herein are methods using particles to clonally amplify templates and to introduce the amplified nucleic acids into many parallel channels with a single template per channel. Once in the channels, the nucleic acids are separated by size using electrophoresis to produce long read length sequencing information. Methods involving optical detection of the size-separated nucleic acids and analysis of the resulting electropherograms to yield the sequences are disclosed.

The invention provides a method for separating high-purity anthocyanin monomers from black rice. The method comprises the following steps of: grinding 5 to 8 percent of episperm on the outer layer of black rice and brown rice to obtain black rice pericarp, leaching anthocyanin in the black rice pericarp by acidized ethanol, and extracting and defatting petroleum ether; performing primary purification of ethanol extracts of the black rice pericarp by using macroporous resin; separating the anthocyanin in coarse extracts of the anthocyanin of the black rice pericarp by the medium pressure preparation liquid phase chromatographic technique, collecting two leading anthocyanin components, which are cyanidin-3-glucoside and paeonidin-3-glucoside, in the black rice pericarp respectively according to the obtained chromatogram, and performing rotary evaporation of eluent at a temperature of below 45 DEG C until all methanol is gone, and performing freeze-drying to obtain powder of the anthocyanin monomers.

The invention relates to a composite solid-phase extraction column used in vegetable pesticide removing. Graphitized carbon black and florisil are filled in a solid-phase extraction hollow pipe, such that the solid-phase extraction column is obtained. A mass ratio of the florisil to the graphitized carbon black is 1:1-5:1. The invention is suitable for normal-phase extractions of low-pole or middle-pole pesticide compounds, and is especially suitable for organic phosphorus pesticide residue removing of colored vegetables and vegetables with relatively complicated groundmass. Compared to other solid-phase extraction columns with single filling materials, the solid-phase extraction column provided by the invention is advantaged in faster and more convenient processing process, large adsorption capacity, and low solvent dosage. The prices of the florisil and the graphitized carbon black are not high. Commercialized solid-phase extraction hollow pipes are adopted. The preparation method is simple, and is suitable for batch productions.

An oligo-proanthocyanidin with high ORAC value and its purification are disclosed. The products contain oligo-proanthocyanidin 45-65wt%, ORAC value is 15000 - 19000 mu-mol TE / g. The process is carried out by dissolving for proanthocyanidin crude product in solvent proportionally, adding into organic feed barrel of organic film ultra-filtration device, ultra-filtering for feed by organic film with molecular weight between 50000-1000000, concentrating and drying to obtain final product. It's convenient and efficient, has more film throughput and yield, and its costs low.

The invention discloses a method for separating and purifying microcystin by utilizing series-connected solid phase extraction columns. The method comprises the following steps of: A, preparing an algae powder material, namely harvesting wild bloom-forming cyanobacteria or microcystis aeruginosa cultured indoors, cooling and drying to prepare dried algae powder; B, preparing the solid phase extraction columns, namely putting C18 fillers into Solid Phase Extraction (SPE) columns; C, extracting algae toxin, namely weighing the dried algae powder, adding methanol according to a proportion, putting on a shaking table and oscillating at room temperature; D, removing impurities, namely adding a supernatant and an organic extractant into a separating funnel according to a proportion, mixing and uniformly shaking, standing, and after delamination, removing the organic extractant; E, separating and purifying the algae toxin, namely after pretreatment, enriching sample solution by using pre-activated series-connected SPE columns, and eluting the toxin by using 20 to 50 percent methanol aqueous solution; and F, performing Methyl Cellulose (MC) detection. The method has good separation effect, large loading amount, is easy and convenient to operate and low in cost, can obtain two microcystins with the weight of more than or equal to 10 mg and higher purity each time, and the High Performance Liquid Chromatography-Ultra Violet / Diode Array Detector (HPLC-UV / DAD) detection can reach 85 to 90 percent.

The invention relates to a method for purifying huperzine A, which is easy for industrialized production. The method comprises the following production steps of: crushing of raw materials, acid water heating and extraction, membrane filtration, impurity removal and condensation, aminated chloroform extraction and condensation, medium-pressure alumina column chromatography, chloroform methanol recrystallization, and drying of a finished product. The method for producing the huperzine A has low energy consumption and short period.

In a spectrophotometer for measuring transmitted light of a trace liquid sample, four sample holders 12 are provided on a disk-like sample plate 11 while spaced apart by 90 degrees. The sample plate 11 is driven to rotate so that each of the sample holders 12 is sequentially moved to a sample supply position U1, a measuring position U2, a wiping position U3 and a waiting position U4. At the sample supply position U1, a trace amount of the liquid sample is dropped into a groove of the sample holder 12. Then at the measuring position U2, a window plate 22 is lowered onto the groove so as to determine the optical path length. Next, measurement of the transmitted light is performed. Further, while the sample holder 12 moves from the measuring position U2 to the waiting position U4, the liquid sample is absorbed and removed by contact with a cleaning pad 26. The liquid sample attached to the window plate 22 is wiped out by another pad. Since the operation to wipe off the measured sample is automatically performed, it is possible to improve the throughput of the measurement.

The invention discloses probes, primers and a kit for detecting a T790M mutation of an EGFR gene. The probes and the primers have the following sequence: SEQ ID NO: 01 to SEQ ID NO. 07. The probes, the primers and the kit have the following benefits: (1) SNP sites on the primers are designed as G / A merged basic groups, so that all efficiencies are compatible, and the amplification efficiency is improved; (2) the sensitivity is high, that is, the detection sensitivity can reach 2 permillage; (3) compared with those adopting a digital PCR method, the operation is simple, the cost is reduced, and the clinical application range is wide; (4) through blood plasma sample detection with a large reaction volume, the DNA loading quantity of blood plasma samples is increased, the detection system is more stable, and the detection rate of the blood plasma samples is improved; (5) the detection speed is high, that is, the detection process can be completed within only 120 minutes, and the time consumed in the detection process is only a half of that consumed according to the digital PCR method; (6) the probes, the primers and the kit, provided by the invention, can be utilized for detecting peripheral blood samples, so that convenient sampling and dynamic detection can be realized.

The present invention separates and purifies taxol by means of column chromatographic process with macroporous resin, such as AB-8, D101, D201 and AK-9 as fixed phase and polar solvent, such as water solution of ethanol or acetone, as flowing phase, with the effluent flow rate is 0.6-1.5 L / hr. The effluent with taxol content lower than cephalotmannine content is concentrated and dried to obtain intermediate taxol product A; and effluent with taxol content higher than cephalotmannine content is concentrated and dried to obtain intermediate taxol product B. The said process can separate and purify coarse taxol product with 9-20% content to obtain intermediate taxol product with 40-60% content in the yield as high as 80%. The process can eliminate water soluble impurity from the coarse product.

The invention relates to a method for preparing 6- gingerol and 8- gingerol with fresh ginger. It is characterized in that it compries following steps: cutting fresh ginger into sheets, drying at lowtemperature, disintegrating, sifting with screen of 100 order, heating and refluxing and extracting with acetic ester or acetone, or extracting through immersion, extracting for three times, combiningextract liquid, decompressing and recovering solvent, getting extract concretem dissloving it with acetic ester, treating it with chromatographic silica gel and washing it with benzinum purificatum acetic ester with gradient solvent, checking washing process by using TLC, decompressing to recover solvent and getting coarse 6- gingerol and 8- gingerol; washing coarse product with chloroform- carbinol solvent in Sephadex LH20 gel column, checking washing process with TLC, and getting washung liquid containing 6- gingerol and 8- gingerol with higher purity, decompressing to recover solvent and getting pure 6- gingerol and 8- gingerol, checking content with HPLC. The invention is characterized by simple operation, high productivity and production of two kinds of product at the same time.

The invention discloses a purification method of insulin determir. The purification method comprises the following steps of: during purification, balancing a cation exchange column by an acidic buffer solution; adjusting the pH value of an insulin determir solution to be less than 5 by the acidic buffer solution or an identically systematic diluted acid, thus obtaining a sample solution; adding the sample solution on the cation exchange column, then washing the cation exchange column by a balanced buffer solution; and successively washing various adsorbed substances down from the column by a 0.5-1.0M acidic buffer solution with a pH value of 2-4 in a liner gradient elution way, and then collecting a main peak, thus obtaining a target product, namely the insulin determir. The purification method not only has the advantages of being convenient to operate, large loading quantity of samples, high recycling efficiency and the like, but also avoids the rigorous requirements that a large quantity of organic reagents and a high pressure chromatographic system are needed in a traditional purification method, so that the purification method is cost-saving, economical and environmentally-friendly, and suitable for industrial production.

The invention discloses a method for separating and purifying microcystin. Said method uses bloom cyanophyte of eutrophic lake as main raw material, and adopts the processes of organic solvent extraction, rapid chromatographic primary separation and C18 reversed phase preparation liquid chromatography so as to implement separation and purification of microcystin MC-RR and [Dha7]MC-RR. Said method adopts isogradient elution, its stability is good, under the condition of preparation chromatography two microcystins can be completely separated. The single sample quantity can be up to 200 micrograms, and the purity of the obtained MC-RR and [Dha7]MC-RR can be detected by using HPLC-UV up to above 95%.

The invention discloses a method for supercritical fluid extraction of waxberry acid A, which comprises the steps of crushing raw materials, adopting supercritical fluid extraction, adding extract into a medium pressure silica gel column for chromatographic purification, performing ethyl acetate-ethanol elution, collecting the waxberry acid A flow, reducing pressure, concentrating and recrystalizing to obtain the waxberry acid A. The preparation method for the supercritical fluid extraction of the waxberry acid A is simple, suitable for large-scale production, low in energy consumption and small in pollution, and is environment-friendly type production process.

The invention relates to a method for the low-pressure silica gel column chromatographic separation of capsaicin and dihydrocapsaicin. Capsaicin and dihydrocapsaicin can be obtained by the following main procedures: using capsaicinoids as a raw material, carrying out low-pressure silica gel column chromatographic separation, eluting with petroleum ether and other solvents, carrying out HPLC (high performance liquid chromatography) analysis and the like. Compared with the traditional method, the invention has the advantages of less environmental pollution, high product purity, low cost and the like, thereby being applicable to industrial production.

The invention relates to a method for preparing high-purity gallnut catechin gallate (GCG). The method comprises the steps of firstly, taking a green tea polyphenol sample, and adding solvent for dissolution; then taking a proper amount of a tea polyphenol dissolving sample, placing the tea polyphenol dissolving sample in a closed container, and conducting high temperature conversion; then taking conversion liquid, conducting separation with a rapid preparative liquid chromatography, and collecting GCG fraction; making the fraction be subjected to evaporation and concentration, so that a GCG monomeric compound crude product is obtained; finally, taking a proper amount of the crude product, and adding solvent for dissolution; conducting separation and purification on the monomeric compound crude product with a high pressure preparative liquid chromatography, and collecting GCG fraction; then making the fraction be subjected to rotary evaporation and concentration, freeze-drying and vacuum drying, so that a GCG monomeric compound is obtained. According to the method, the high temperature conversion method is adopted, and the content of gallnut catechin gallate (GCG) in a green tea polyphenol solution is raised.

The invention relates to a method for separating and refining arteannuin, dihydro-artemisinic acid and artemisinic acid by reversed-phase high performance liquid chromatography. The method mainly comprises the following steps of: separating and purifying an artemisia apiacea crude extract extracted by organic solvent by reversed-phase high performance liquid chromatography, and concentrating and crystallizing to obtain pure products of the arteannuin, the dihydro-artemisinic acid and the artemisinic acid. The method is high in yield, separation efficiency and a separation speed, the pure products of the arteannuin, the dihydro-artemisinic acid and the artemisinic acid can be obtained simultaneously, and the purity can reach over 96 percent, so the method can be used for large-scale industrial production.

A gene chip surface processing method includes: pretreating slide by siliconizing reagent, acrylic acid and acrylic acid monomer polymerizing on slide surface by polymer polyreacting, forming covalent connecting, inducing active molecule when polyreacting, chemical reacting with amide group of polymer, and forming amino-sensitive chemical surface. It achieves low cost, simple process, and good stability.

The invention discloses a method for preparing a saliva protein sample, and belongs to the technical field of preparation of saliva protein samples. The method is characterized by comprising the following steps of: 1) collecting saliva; 2) centrifuging 800g of collected saliva at 4 DEG C for 15 minutes, and sucking supernate; 3) adding 0.05 to 5 volume percent acetic acid and uniformly mixing; 4) centrifuging 14,000g of the mixed solution at 4 DEG C for 60 minutes, and sucking supernate; 5) putting the supernate in a dialysis bag, and dialyzing by using double distilled water for 5 hours; and 6) condensing to obtain the saliva protein sample. The method has the advantages of high protein recovery rate, short preparation time, safety, nontoxicity, increase of loading quantity of low abundance proteins, clear and easily analyzed maps and the like.

The invention discloses a method for extracting erianin from dendrobe. The method comprises the following steps: extracting by using an alkaline organic solvent, mixing and extracting by a nonaqueous phase organic solvent, performing small-pore diameter resin separation, crystallizing by organic solvents, and preparing to obtain high-purity erianin square crystal. Compared with a conventional process method, the method has the advantages that the process is simple, the crude extraction solvent is low in cost, the target is fewer in non-aqueous phase extraction impurities, the column chromatography cost is low, the packing is high in separation precision and can be repeatedly utilized, the chromatography solvent cost is low, the environmental pollution is relatively low, the erianin is high in yield, the purity is high and the like. Moreover, the method is suitable for industrial production.

The invention discloses a cloprostenol sodium pure product preparation method for preparation of cloprostenol sodium by an industrial preparative liquid chromatography separation purification system, and belongs to the technical field of separation and purification of biological products, the method is simple and high in yield, the prepared cloprostenol sodium is high in product purity, and the method is applicable to industrial mass production preparation of the cloprostenol sodium.

The invention relates to a method for separating III type precollagen amino terminal peptides, which solves the problems of an extraction method, the efficiency, the purity and the activity of the III type precollagen amino terminal peptides. An ammonia sulfate fractionation method is adopted, so that the quantity of impurities is reduced by 5.5 percent, and the yield is increased by 11.7 percent in a first step; Qsepharose FF (Qsepharose Fast Flow) is adopted, so that the loading amount is large, the flow rate is high, and further separation is achieved; a buffer solution with the pH value of 4.5 and a buffer solution with the pH value of 8.5 are not used for eluting alternatively, but hydrophobic chromatography which plays a role in protecting proteins is adopted, so that purity and activity are greatly enhanced; Sephacryl 200 is replaced by high-resolution Superdex 200, so that the purity is further enhanced; and by SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis) analysis, the purity is up to 98 percent, and the activity is 10 times that of the prior art.

The invention discloses a method for preparing przewaquinone sodium sulfonate. The preparation method comprises the following steps: separating tanshinone sulfonated liquid by adopting high performance liquid chromatography, wherein the conditions of the high performance liquid chromatography are as follows: the mobile phase A is 0.1% triethylamine-water, the mobile phase B is 0.1% triethylamine-methanol, or the mobile phase A is 0.1% diethylamine-water, the mobile phase B is 0.1% diethylamine-methanol, or the mobile phase A is 0.1% ethylamine-water, the mobile phase B is 0.1% ethylamine-methanol, or the mobile phase A is water, the mobile phase B is methanol, the percent is the volume percent of the component in the mobile phase, and the detection wavelength is 271nm. The preparation method disclosed by the invention is strong in preparation ability, large in preparation quantity, short in cycle, and high in efficiency, an organic solvent is reduced, the process operation continuity is strong, and mass control and production are easily carried out.

The present invention pertains to the field of biopharmaceutical processes, more specifically relates to a method to increase salt ion concentration of a buffer to purify a virus, and aims at the problems that virus purifying methods in the prior art are high in cost and not suitable for industrialized production, and the method comprises the following steps: A, inoculation and culture of host cells; B, amplification and lysis of the virus-infected host cells to obtain virus-containing lysate; C, concentration of the lysate to obtain concentrated lysate; D, use of a high salt ion buffer to filter the concentrated lysate, and use of ion exchange chromatography for purifying to obtain the purified virus. The method can obviously remove a large number of impure proteins such as serum and hostcell proteins, increases the sample loading size of the chromatography, and increases the carrying capacity of packing. The method is simple, economical and efficient, is suitable for industrialization and has a good application prospect.

The invention belongs to the technical field of microbial extraction, and relates to equipment and a method for extracting vitamin K2 from microbial fermentation liquor. The equipment comprises a continuous dynamic countercurrent extractor and a medium-pressure preparative chromatography, the medium-pressure preparative chromatography comprises at least one chromatographic group, each chromatographic group comprises four chromatographic columns connected in parallel, each chromatographic column is independently provided with a feed port, an eluent inlet A, an eluent inlet B and an eluent inletC, the feed port is connected with an extracting solution outlet of the continuous dynamic countercurrent extractor, the eluent inlet A, the eluent inlet B and the eluent inlet C are respectively connected with an eluent source A, an eluent source B and an eluent source C, the opening and closing of the four material inlets are respectively and independently controlled by valves, and the valves of different chromatographic columns in the same chromatographic group are synchronously switched. By adopting the equipment provided by the invention, the purity and yield of the vitamin K2 can be improved, and the whole extraction process can completely realize continuous automatic operation.

The invention discloses a manufacturing technique for extracting, separating and purifying kilogram-grade scale high purity GM1 using fresh pig brain tissue as raw material, which is characterized in that large sized extraction pot and adsorption resin with large holes, DEAE Sephadex A-25 and silica gel layer are utilized to build a set of manufacturing technique. The invention has the advantagesof elimination of toxic solvent chloroform in mobile phase in traditional method, less pollution for whole process, simple process, low cost and applicability to industrialized production.

The invention discloses a method for extracting suaveolic acid A. The method is characterized by comprising steps as follows: getting the aboveground part of head-shaped suaveolens as the raw material; crushing the tissue, and carrying out enzymolysis by virtue of cellulose, so as to obtain the raw material subjected to enzymolysis; carrying out supercritical CO2 extraction; separating and collecting to obtain triterpenes extractive; then purifying by high-pressure silica gelcolumn chromatography; collecting fraction of the suaveolic acid A; concentrating in a pressure reducing way; and recrystallizing to obtain the suaveolic acid A. The method disclosed by the invention is simple in operation, suitable for large-scale production, low in energy consumption, low in pollution and short in production period, and is an environment-friendly production technology.

The invention relates to a preparation method of high-purity paeoniflorin and albiflorin. The method includes the steps of firstly, adding smashed paeoniaceae plant raw materials containing paeoniflorin and albiflorin to an ethanol solution, conducting ultrasonic extraction, concentrating an extracting solution to obtain extract, dissolving extract with water, and conducting extraction and pressure reduction concentration to obtain extract; secondly, eluting extract with an alcohol-water solution through column chromatography with macroporous resin as filler, collecting flow components containing paeoniflorin and albiflorin, and conducting concentrating and drying to obtain coarse extract; thirdly, separating out the coarse extract through a high-speed countercurrent chromatography method, conducting online monitoring through an ultraviolet detector, collecting flow components and pressure reduction concentration, and conducting crystallizing and drying to obtain paeoniflorin and albiflorin, wherein a solvent system of the high-speed countercurrent chromatography is prepared from carbon trichloride, butyl alcohol, methyl alcohol and water. Paeoniflorin and albiflorin prepared through the method are high in product purity and good in quality, are suitable for high-speed countercurrent chromatographs of various types and can be easily and industrially amplified.

The invention discloses a thunberginol E extraction method. The method is characterized in comprising the following steps: hydrangea macrophylla leaf is adopted as a raw material; through tissue crushing and cellulase enzymolysis, an enzymolyzed raw material is obtained; the material is added into a flash extractor and is subjected to flash extraction; filtering is carried out, such that an extract liquid is obtained; concentration and drying are carried out; high-pressure silica gel column chromatographic purification is carried out; thunberginol E fraction is collected; reduced-pressure concentration is carried out; and acetone recrystallization is carried out, such that thunberginol E is obtained. The method provided by the invention is simple to operate, and is suitable for large-scale productions. The method has the advantages of low energy consumption, low pollution, and short production period. The method is an environment-friendly production process.

The invention discloses a purifying method for nostoc sphaeroids kutzing phycobiliprotein. The purifying method comprises the steps: (1) choosing an anion exchange column to perform column chromatography; (2) preparing a PB buffer solution Buffer A and a PB-NaCl buffer solution Buffer B; (3) dissolving nostoc sphaeroids kutzing phycobiliprotein coarse extract powder through the PB buffer solution Buffer A, centrifuging and filtering; (4) washing a pump and utilizing the PB buffer solution Buffer A to balance an ion exchange column; (5) directly loading sample of the nostoc sphaeroids kutzing phycobiliprotein which is prepared in the step (3) and dissolved into the PB buffer solution Buffer A, performing gradient elution through the PB-NaCl buffer solution Buffer B and collecting eluant; (6) utilizing a saline solution to clean the iron exchange column and reservedly cleaningback flushing through an alkali solution; (7) performing ultrafiltration and concentration on leacheate and dialyzing to obtain a purified nostoc sphaeroids kutzing phycobiliprotein water solution. The method disclosed by the invention is simple to operate, the concentration and the purity of the obtained nostoc sphaeroids kutzing phycobiliprotein are both obviously improved, and the method is suitable for industrial large-scale production.

The invention relates to a preparation method of high-purity paeoniflorin and albiflorin. The method includes the steps of firstly, adding smashed paeoniaceae plant raw materials containing paeoniflorin and albiflorin to an ethanol solution, conducting ultrasonic extraction, concentrating an extracting solution to obtain extract, dissolving extract with water, and conducting extraction and pressure reduction concentration to obtain extract; secondly, eluting extract with an alcohol-water solution through column chromatography with macroporous resin as filler, collecting flow components containing paeoniflorin and albiflorin, and conducting concentrating and drying to obtain coarse extract; thirdly, separating out the coarse extract through a high-speed countercurrent chromatography method, conducting online monitoring through an ultraviolet detector, collecting flow components and pressure reduction concentration, and conducting crystallizing and drying to obtain paeoniflorin and albiflorin, wherein a solvent system of the high-speed countercurrent chromatography is prepared from carbon trichloride, butyl alcohol, methyl alcohol and water. Paeoniflorin and albiflorin prepared through the method are high in product purity and good in quality, are suitable for high-speed countercurrent chromatographs of various types and can be easily and industrially amplified.