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 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.

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 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 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 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 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 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 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.