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45 results about "Resazurin" patented technology

Resazurin (7-Hydroxy-3H-phenoxazin-3-one 10-oxide) is a blue dye, itself weakly fluorescent until it is irreversibly reduced to the pink colored and highly red fluorescent resorufin. It is used as an oxidation-reduction indicator in cell viability assays for both aerobic and anaerobic respiration. Usually it is available commercially as the sodium salt.

Novel strain for producing butanol and method for producing butanol

The invention discloses a novel clostridium beijerinckii strain for producing butanol. The strain has a classification name of clostridium beijerinckii IT66 (Clostridium beijerinckii IT66) and has a collection number of CCTCCM NO:2011404. The invention also discloses application of the clostridium beijerinckii in fermentation production of the butanol. The clostridium beijerinckii IB4 is subjected to continuous culture and plasma induced mutation by adopting a chemostat, and a strain which tolerates an inhibitor and is high in reducing power is screened through a resazurin flat plate and is high in acidolysis byproduct resistance, high in butanol ratio and high in solvent yield. When a non-detoxified corncob acidolysis sugar solution is a carbon source, the total solvent yield and butanol yield in a 5L fermentation tank respectively reach 11.3g / L and 9.1g / L and are respectively improved by 2.3 times and 2.4 times compared with those of an original strain cultured under the same conditions, the butanol ratio is 80 percent, the sugar conversion rate is 0.32, and the strain has significant social meaning and economic value.
Owner:NANJING UNIV OF TECH

Separation purification process and application of anaerobic degradation pure culture of polycyclic aromatic hydrocarbon

InactiveCN101195811AGood anaerobic degradation performanceBacteriaContaminated soil reclamationVitamin CPurification methods
The invention discloses a separated purification method and the application thereof of a polycyclic aromatic hydrocarbon anaerobic degrading pure strain, which belongs to the polluted soil and aqueous biological degradation handling field. A base bottom layer flat plate is manufactured in the a anaerobic box filled with nitrogen gas by taking polycyclic aromatic hydrocarbon as a sole carbon source and an energy source and taking sodium nitrate or potassium nitrate as an electronic accepted anaerobic agar solid culture, the top layer flat plate of the anaerobic agar solid culture including polycyclic aromatic hydrocarbon anaerobic degrading mixed bacteria is manufactured on the bottom layer flat plate, the manufactured culture medium flat plate is put in a drying container filled with the nitrogen gas to culture one to two months after being sealed. Polycyclic aromatic hydrocarbon, electronic acceptance, basic culture medium, trace metal liquid, vitamin c solution and colony in the culture medium flat plate connected into the anaerobic bottle of resazurin are added in the anaerobic box filled with the nitrogen gas, and are cultured for 20 to 25 days in an oscillator under the temperature of 20 plus or minus 2 DEG C and the rotating speed of 100 r / min. Then, the cyclical switchover is performed, after a cycle time is larger than four times, the pure strain of the anaerobic degrading polycyclic aromatic hydrocarbon can be obtained. The method can obtain good microbe pure strain to the polycyclic aromatic hydrocarbon anaerobic degrading performance by being separated.
Owner:BEIJING NORMAL UNIVERSITY

Method for rapidly screening and evaluating antibiotic bioactivator

The invention discloses a method for rapidly screening and evaluating an antibiotic bioactivator, in particular a test method for screening the antibiotic bioactivator by combining an accelerated solvent extraction apparatus and a microplate resazurin assay. The method comprises the following steps of: preparing a bioactivator by the accelerated solvent extraction apparatus; adding the bioactivator and known antibiotic into a microplate according to concentration gradient, and adding experimental bacterial suspension into various holes; placing the microplate under suitable conditions, incubating, adding resazurin solution, and mixing uniformly; determining light absorption values of the holes of the microplate at 600nm by using a microplate reader; evaluating a minimum inhibitory concentration (MIC) value of the bioactivator to be screened according to light absorption values in known antibiotic holes and light absorption values in bioactivator holes at different concentrations; and further evaluating the antibiotic activity of the bioactivator. The method is rapid, simple and convenient, and the antibiotic activity of the bioactivator can be objectively and accurately evaluated, so that the high-throughput screening of the antibiotic bioactivator becomes possible.
Owner:HAINAN UNIVERSITY

Mycobacterium tuberculosis medicament sensitive phenotype detection method and application of method

The invention discloses a mycobacterium tuberculosis medicament sensitive phenotype detection method, which comprises the following steps of: 1) adding 2.5 to 6mul of 0.2 percent of resazurin dissolved by using methanol with mass percentage concentration into the inner side of a tube cover of a sterile centrifuge tube, and sterilizing the centrifuge tube after the methanol is volatilized; 2) inoculating 5 to 50mg of mycobacterium tuberculosis to be detected into a 7H9-S culture medium, regulating the concentration of the bacteria solution by adopting turbidimetry, regulating the concentration of the bacteria solution to the concentration of a Mcburney turbidimetric 1 tube by using sterile water or 0.9-percent physiological saline, and diluting the bacteria solution by 20 times by using the 7H9-S culture medium; 3) inoculating 50 to 150mu of diluted bacteria solution into the centrifuge tube of the step 1); 4) adding 100mul of anti-tubercular medicament solution diluted by 7H9-S and to be detected into the centrifuge tube of the step 3), wherein the concentration of the medicament to be detected is 0.01mug / ml to 2.5mg / ml; 5) after the centrifuge tube of the step 4) is covered closely, culturing the solution for 6 to 7 days at the temperature of 37 DEG C; and 6) inverting and oscillating the centrifuge tube of the step 5), and culturing the solution for 12 to 48 hours at the temperature of 37 DEG C after the solution becomes blue. Proved by experiments, in the mycobacterium tuberculosis medicament sensitive phenotype detection method and application of the mycobacterium tuberculosis medicament sensitive phenotype detection method in anti-tubercular medicament screening, the method has low workload and short detection time; compared with a traditional L-J medium medicament sensitive detection method, the accuracy reaches 90 to 99 percent; and the method has good biological safety, does not cause laboratory propagation of tuberculosis, is suitable for large-scale.
Owner:LANZHOU UNIVERSITY

High-throughput rapid antimicrobial susceptibility test kit for veterinary clinical application

The invention discloses a high-throughput rapid antimicrobial susceptibility test kit for veterinary clinical application and a method. The kit comprises an independently-packaged enrichment medium, an antimicrobial susceptibility test plate and reaction liquid; the enrichment medium is selected from a sterile 1640 medium or an MH medium with 1% sterile neonatal bovine serum and 0.1% sterile NAD added; the antimicrobial susceptibility test plate is a reaction plate coated with a plurality of antibiotics, each hole of the reaction plate is coated with an antibiotic, and the concentration of theantibiotics is 1-2 times of the maximum plasma concentration after the clinical administration of the antibiotics; and the reaction liquid is a 0.2% resazurin sterile solution. According to the kit and the method, an antimicrobial susceptibility test is carried out, the time for the result is short, more drugs are tested at a time, the kit and the method are easy to operate, and special instruments and equipment are not needed; the test process is a liquid-phase reaction, more accords with the physiological environment and is close to the actual production; and the result is visual, specifically, blue represents susceptible, red represents susceptible, and the closer the color is to blue, the more susceptible a drug is.
Owner:苏州艾可瑞动物检测技术服务有限公司 +2

Enrichment culturing method for anaerobic microorganisms

The invention discloses an enrichment culturing method for anaerobic microorganisms. The method comprises the steps that after a culture medium is prepared, resazurin is added to serve as an oxidation-reduction indicator; the culture medium is put into a high-pressure sterilizing pot to be sterilized for 20-30 min at 121 DEG C and then taken out to be put into an ultra-clean bench to be cooled to 40 DEG C-60 DEG C for standby application; a deoxidant L-cysteine and sodium salt of the L-cysteine are added in the processed culture medium; a normal saline solution in a normal saline injection bottle is completely drawn with an injector needle, the normal saline injection bottle is cleaned with sterile water for 4-6 times and then placed on the ultra-clean bench, ultraviolet sterilizing is performed for 20-30 min, the culture medium and an inoculating sample are injected into the injection bottle, air in the bottle is completely drawn with an injector, the bottle is filled with high-purity nitrogen, the normal saline bottle is placed in a constant-temperature culturing box to be cultured for 2-5 days at 37 DEG C, and then enrichment culturing of the anaerobic microorganisms is completed. The enrichment culturing method for the anaerobic microorganisms does not need the help of an anaerobic culturing box and is convenient, fast and low in cost, and sterility in the whole course can be guaranteed.
Owner:GUILIN UNIVERSITY OF TECHNOLOGY

Sulfated bile acid enzyme fluorescence capillary analytical method and enzyme fluorescence quantitative reagent kit

The invention discloses a sulfated bile acid (SBA) enzyme fluorescence capillary analytical method and an SBA fluorescence fixed amount agent kit (SBA-FCA-Kit) which belongs to the field of a clinic biochemistry inspection field and is suitalbe for the fast screening and diagnosing of liver and gall diseases, more particularly, is suitalbe for the early period discovery of jaundice in neonatal period and congenital biliary atresia. After being mixed with fluorescence reaction liquid, a biology sample is sucked by an SBA capillary biology reactor (SBA-CBR) which contains sulfated bile acid sulfoacid esterase and Beta-hydroxysteroid dehydrogenase to determine the empty fluorescence intensity of the SBA-CBR under a given excitation wavelength and an emission wavelength; then the biology sample is led to continuously react for a certain time and the fluorescence intensity is determined; the amount of the SBA in the sample is fixed by using fluorescence intensity difference value. The SBA-FCA-Kit is composed of an SBA-CBR and fluorescence reaction liquid containing Beta-NAD<+>, an electric transmission body and a diazoresorcinal, etc.
Owner:SICHUAN UNIV

Method for producing hydrogen through intensified anaerobic fermentation

InactiveCN102876723AImprove permeabilityOvercome the high cost and low efficiency of hydrogen productionFermentationNitrogenResazurin
The invention discloses a method for producing hydrogen through intensified anaerobic fermentation. The method comprises the following steps of: adding a culture medium, resazurin and a cetyl trimethyl ammonium bromide solution into a fermentation container, mixing uniformly, sterilizing, inoculating liquid obtained after cow dung is fermented, introducing sterile nitrogen into the fermentation container and an alkaline washing bottle which is connected with the fermentation container so as to displace the air, sealing a sterile nitrogen introduction port, performing shake culture at constant temperature, performing alkaline washing on gas produced by fermentation, and collecting to obtain the hydrogen. By adding cetyl trimethyl ammonium bromide into a fermentation substrate, the problems that the cost of the current hydrogen production by biological anaerobic fermentation is still high and the hydrogen production efficiency is low are solved, and the effects that the accumulated hydrogen yield is improved by 38 percent and the maximum hydrogen production rate is improved by 44 percent are successfully achieved. The method can be simply operated, and is low in cost and easy to implement.
Owner:HENAN UNIV OF SCI & TECH

Method for producing hydrogen by fermenting special anaerobic clostridium butyricum

The invention provides a method for producing hydrogen by fermenting special anaerobic clostridium butyricum. The screened special anaerobic bacterium is clostridium butyricum JCM1391. Proven by tests, the hydrogen is produced by fermenting by batch by using anaerobic bacteria under the sterile anaerobic condition, wherein the mixture of 10.0-15.0g of peptone, 2.0-3.0g of beef extract, 5.0-10.0g of yeast extract, 3.0-4.0g of Na2HPO4, 0.2-0.5g of L-cysteine, 0.2-0.5g of L-cysteine hydrochloride monohydrate, 1 ml of resazurin indicator which accounts for 0.2 percent by weight and 1 ml of distilled water is used as culture media, and glucose is used as fermentation matrix. The method has the advantage of high hydrogen production efficiency and low requirement on environmental condition for fermentation.
Owner:SOUTHEAST UNIV

Clostridium acetobutylicum strain and screening method and application thereof

The invention relates to a Clostridium acetobutylicum strain capable of quickly generating butanol by fermenting xylose and starch, and a screening method and application thereof. The invention discloses Clostridium acetobutylicum, which is classified and named Clostridium acetobutylicumXY16 with the preservation and registration number of CCTCC NO:M2010011. The strain obtained by ion beam mutation and screening of a xylose plate, a 2-deoxy-D-glucose plate, a bromocresol green plate and a resazurin plate can quickly and efficiently generate the butanol by fermenting xylose and starch, has the advantages of high yield of total solvent, high butanol ratio and good repeatability, and is a good strain suitable for industrial production.
Owner:NANJING UNIV OF TECH

Sulfated bile acid enzyme fluorescence capillary analytical method and enzyme fluorescence quantitative reagent kit

The invention discloses a sulfated bile acid (SBA) enzyme fluorescence capillary analysis and an SBA fluorescence quantitative test kit (SBA-FCA-Kit), which belongs to the field of a clinic biochemistry inspection field and is suitable for the fast screening and diagnosing of liver and gall diseases, more particularly, is suitable for the early period discovery of jaundice in neonatal period and congenital biliary atresia. After being mixed with fluorescence reaction liquid, a biology sample is sucked by an SBA capillary biology reactor (SBA-CBR) which contains sulfated bile acid sulfoacid esterase and Beta-hydroxysteroid dehydrogenase to determine the blank fluorescence intensity of the SBA-CBR under a given excitation wavelength and an emission wavelength; then the biology sample is ledto continuously react for a certain time and the fluorescence intensity is determined; the amount of the SBA in the sample is determined by using fluorescence intensity difference value. The SBA-FCA-Kit is composed of an SBA-CBR and fluorescence reaction liquid containing Beta-NAD<+>, an electric transmission body and diazoresorcinal, etc.
Owner:SICHUAN UNIV

Bergenia crassifolia extract and extracting method and medical application in oral diseases

The invention provides a bergenia crassifolia extract and an extracting method and medical application in oral diseases. Through a resazurin dyeing method and an XTT method, the extract of bergenia crassifolia is subjected to bacteriostatic activity and inhibitory biological film activity study, the extract of bergenia crassifolia has an obvious inhibitory effect on streptococcus mutans, streptococcus sanguis, streptococcus salivarius, lactobacillus acidophilus, porphyromonas gingivalis, fusobacterium nucleatum, actinobacillus actinomycetemcomitan and biological film of the fungi, has a better inhibitory effect on streptococcus mutans, porphyromonas gingivalis and the biological film of the fungi and can be used for preparing daily necessities and medicine for preventing and treating the oral diseases.
Owner:NORTHEAST NORMAL UNIVERSITY

Method for sifting or detecting antibiotic active compound from plant extracts

The invention discloses a method for screening or detecting antibacterial active substance from plant extract. The method includes the following steps: (1) plant extract is prepared, or an extract bank is created based on the plant extract, or constituent or ingredient is obtained through separation and purification; (2) cultivation is completed according to the culture conditions of different bacteria and fungi; (3) each sample to be tested is added in bacterium culture solution or fungus culture solution; (4) 12 hours to 24 hours cultivation is completed under bacterium culture condition or fungus culture condition; (5) resazurin indicator is added in each culture solution; (6) according to the color change of resazurin, bacterium survival status is judged so as to judge whether the constituent or ingredient of the plant extract has bacteriostatic activity. With easy operation, simple experiment apparatus and less work load, the invention can obtain obvious results; moreover, the invention realizes fast screening of plant extract with antibacterial activity and screens out antibacterial active constituent or ingredient from the plant extract quickly, thereby providing basis for drug research; in addition, the invention can also be used in detecting plant antibacterial active constituent.
Owner:GUIZHOU UNIV

Method for detecting D-2-hydroxyglutarate by using FAD dependent D-2-hydroxyglutarate dehydrogenase and resazurin

The invention discloses a method for detecting D-2-hydroxyglutarate by using FAD dependent D-2-hydroxyglutarate dehydrogenase and resazurin, and relates to the technical field of enzymatic detection.The method uses FAD dependent D-2-hydroxyglutarate dehydrogenase, resazurin and a PBS buffer solution to realize detection of D-2-hydroxyglutarate; and the FAD dependent D-2-hydroxyglutarate dehydrogenase is an AX-F-D2HGDH protein or a P-D2HGDH protein. The method is simpler in composition, lower in cost and simpler and more convenient to operate. Since no cofactor, coenzyme or diaphorase needs tobe added, introduced interference is less, sensitivity is higher, only one enzyme needs to be prepared, no exogenous cofactor NAD and the like need to be added, and meanwhile, the buffer solution canbe a PBS buffer solution which is lower in cost and easy to obtain, so that cost is lower.
Owner:THE SECOND HOSPITAL OF SHANDONG UNIV

Display experiment method and device for coupling internal flow of bubbles and external mass transfer of bubbles

ActiveCN113237793AShow flow phenomenonShows extravesicular mass transferSurface/boundary effectMaterial analysis by optical meansImage manipulationEngineering
The invention relates to a display experiment method and device for coupling bubble internal flow and bubble external mass transfer. The method comprises the following steps of adding a prepared resazurin solution with a certain concentration, a sodium hydroxide solution and a glucose solution to a certain liquid level in a bubble observation chamber by utilizing a resazurin chromogenic reaction and taking smoke as tracer particles, introducing a mixed gas of oxygen and smoke, and by virtue of a high-speed camera and an image processing system, meanwhile, the phenomena of fluid flowing inside the bubbles and liquid phase mass transfer outside the bubbles being displayed. The method is advantaged in that an internal flow phenomenon of the captured bubbles is obvious, and the mass transfer wake outside the bubbles is clear.
Owner:QINGDAO UNIV OF SCI & TECH

Pure culture method of anaerobic microorganisms

The invention discloses a pure culture method of anaerobic microorganisms. The pure culture method comprises the following steps of: step 1, preparing a liquid culture solution and boiling; dropwise adding resazurin; sub-packaging into a thin-opening pipe to obtain a culture pipe; step 2, introducing N2 / H2 mixed gas and aerating; rapidly covering a rubber plug tightly; step 3, putting the culture pipe into a high-pressure sterilization pot and sterilizing at high temperature; transferring into a super-clean platform and carrying out ultraviolet sterilization; and step 4, injecting a magnetic antioxidant into the culture pipe by utilizing an injector and uniformly mixing; injecting an anaerobic microorganism seed solution into the culture pipe by utilizing the injector; putting the culture pipe into a constant-temperature culture box and culturing to obtain an anaerobic microorganism group. The pure culture method of the anaerobic microorganisms has the beneficial effects that (1) an anaerobic culture box does not need to be utilized and the operation is simple and convenient; and (2) the magnetic antioxidant is not dissolved into the liquid culture solution and is not easily absorbed by the anaerobic microorganisms, so that the growth is not influenced; the separation of the anaerobic microorganisms can be simplified through magnetic separation and the relatively pure anaerobic microorganism group is obtained.
Owner:DONGGUAN UNIV OF TECH

Culture medium and preparation method thereof

The invention provides a culture medium. The culture medium is prepared from a bacteriostatic agent which is prepared from salt and anthocyanin. The culture medium does not contain antibiotics, the antibiotics is replaced by the anthocyanin to play role of inhibiting miscellaneous bacteria. Meanwhile, the effect of indicators is achieved by ingeniously utilizing the phenomenon that the anthocyaninproduces acid along with lactic acid bacteria to change color, the indicators such as resazurin, bromcresol purple, bromocresol green, litmus, MTT and TTC are avoided, purple potato starch can promote proliferation of bifidobacteria, and then the bifidobacteria can be easily and safely separated out from feces.
Owner:INNER MONGOLIA MENGNIU DAIRY IND (GRP) CO LTD

Method for detecting fluorescence or absorbance, method for suppressing background, method for measuring ADP, method for measuring activity of ADP-synthesizing enzyme, and method for measuring activity of glucosyltransferase

A method for detecting fluorescence or absorbance according to the present invention is characterized by comprising reducing resazurin into resorufin by a diaphorase in the presence of an SH reagent and NADH or NADPH and then measuring the resultant fluorescence intensity or absorbance. A method for measuring ADP according to the present invention is characterized by comprising: step (2-1) for treating glucose with ADP and ADP-dependent hexokinase; step (2-2) for treating glucose-6-phosphate obtained in the above step (2-1) with NAD or NADP and glucose-6-phosphate dehydrogenase; and step (2-3) for treating resazurin with NADH or NADPH obtained in the above step (2-2) and a diaphorase in the presence of an SH reagent and then measuring the resultant fluorescence intensity or absorbance.
Owner:THE UNIV OF TOKYO

Method for rapidly identifying anaerobic microorganisms in wine brewing

The invention discloses a method for rapidly identifying anaerobic microorganisms in wine brewing, and belongs to the technical field of anaerobic microorganism identification. The method for rapidlyidentifying the anaerobic microorganisms in the wine brewing comprises the following steps: S1, preparing an anaerobic culture solution, namely uniformly mixing 0.18 g of potassium persulfate, 0.2993g of potassium phosphate, 0.35 g of sodium chloride, 0.40 g of ammonium persulphate, 0.040 g of magnesium chloride, 0.085 g of potassium sulfate, 0.002 g of resazurin, 0.6 g of L-cysteine, 0.7 g of L-ascorbic acid, 5.0 g of sodium bicarbonate, 1.3 g of peptone and 1500 mL of distilled water so as to obtain an anaerobic culture solution for standby application; S2, preparing a bacterial suspension,namely weighing 15 g of fermented grains in the presence of a nitrogen flow, adding the weighed fermented grains into 95 mL of sterilized normal saline in an anaerobic bottle, quickly and tightly covering the anaerobic bottle with a cap, oscillating the anaerobic bottle on a shaking table at a temperature of 50 DEG C and a rate of 200 r / min for 15 min, taking the oscillated anaerobic bottle out,putting the anaerobic bottle into an anaerobic workstation, sucking 0.2 mL of liquid supernatant, and adding the liquid supernatant into the sterilized culture solution so as to carry out anaerobic accumulation culture for 36 hours; S3, carrying out colony culture; and S5, carrying out identification. The method for rapidly identifying the anaerobic microorganisms in the wine brewing is convenientto operate, simple in processm, and hign in identification accuracy.
Owner:樊海麟

Intestinal anaerobic microorganism culture bottle and preparation method thereof

The invention discloses an intestinal anaerobic microorganism culture bottle and a preparation method thereof. The culture bottle comprises a culture bottle sealed and packaged and a culture solutionfilled in the culture bottle. The prepration method comprises the steps of sequentially mixing a basic culture medium solution, an L-cysteine saline solution and a resazurin aqueous solution in proportion to obtain a basic culture solution, filling the culture bottle with the basic culture solution, and sealing and packaging the culture bottle; performing high-pressure sterilization on the sealedculture bottle; and finally, mixing a 1640 culture medium solution, BI serum, vitamin K and hemin in proportion to obtain an active additive mixed solution, injecting the active additive mixed solution into the culture bottle which is subjected to high-pressure sterilization and cooled to room temperature, thereby completing the preparation. According to the anaerobic microorganism culture bottle,a culture bottle body and a packaging piece are adopted, all components for preparing the culture solution are commercially available products, the raw materials are easy to obtain, the threshold andcost of anaerobic culture are reduced, anaerobic culture, storage and transportation are easier to operate, the preparation method is simple and easy to implement, the requirement for large-scale production is met, and the use requirements of hospitals and scientific research institutions are met.
Owner:TIANJIN MEDICAL UNIV

Strain preservation method of anaerobic methanogens in coal seams

The invention belongs to the technical field of microbial engineering. In order to solve the problems that the existing strain preservation method cannot meet the requirements of short-term and long-term preservation of strains of methanogens produced by coal-bed methane and cannot maintain the activity of high-yield methane gas of the strains, the invention provides a strain preservation method of anaerobic methanogens in coal seams. The method comprises the steps of screening a strain to be preserved, adding the screened strain into a 5ml of anaerobic tube, then adding an equivoluminal sterilized glycerin / deoxidized L-cysteine solution, evenly mixing the mixed solution, and then preserving; long-term preservation: mixing lump anthracite coal, a preservation nutrient solution, L-cysteine, resazurin and a bacterium solution according to a certain ratio and then preserving the mixture at room temperature, simulating the environmental conditions of the coal seams, and enabling the methanogens to maintain higher activity and a more stable bacterial flora structure all the time so as to obtain more ideal methane yield. The strain preservation method does not need ultra-low temperature equipment and the like, does not need special equipment, is convenient and fast, is suitable for short-term and long-term preservation of different amounts of strains, is economical and convenient, and lowers the production and test costs.
Owner:SHANXI JINCHENG ANTHRACITE COAL MINING GRP CO LTD

Method for detecting PBMC activity under in-vitro 3D culture condition by using resazurin reagent

The invention discloses a method for detecting PBMC activity under an in-vitro 3D culture condition by using a resazurin reagent. The method comprises the following specific steps: step 1, preparing resazurin mother liquor; step 2, preparing a resazurin detection reagent; and step 3, determining the activity of PBMC under an in-vitro 3D culture condition. The standard resazurin solution which canbe produced on a large scale and is used for detecting the activity of human peripheral blood mononuclear cells after being cultured in an in-vitro 3D culture system in vitro is established; the reagent is high in cell activity detection sensitivity, convenient to operate, free of adverse effects on cells and free of pollution to the environment, and it is proved through actual tests that the reagent is suitable for detecting the activity of PBMC in a 3D culture system.
Owner:郑州鲲鹏健康科技有限公司

Method for quickly screening anti-Aspergillus-niger active substances

The invention relates to a method for quickly screening anti-Aspergillus-niger active substances, which comprises the following steps: adding a sample to be screened, a resazurin and Aspergillus niger spore bacterial suspension, into a 96 microporous plate, and culturing in a 37 DEG C constant-temperature culture box for some time to obtain the result, wherein a negative control and a positive control are arranged, and the preferable culture time is 12-36 hours; and screening to obtain the anti-Aspergillus-niger active substances according to the result. When the color of the microporous plate is blue or purple, the fluorescent value measured by the microplate reader is lower than the 15000; and when the inhibition rate is higher than 40%, the detected sample is the anti-Aspergillus-niger active substances. The method is simple, quick and accurate.
Owner:THIRD INST OF OCEANOGRAPHY STATE OCEANIC ADMINISTATION

Deoxidizing method of fluid medium

InactiveCN107904176AAvoid poisoningSmooth enrichmentMicroorganismsPink colorFree state
The invention provides a deoxidizing method of a fluid medium. The method comprises the following steps: S1, preparing a resazurin water solution to serve as an anaerobic indicator which takes on a dark blue color or a light blue color in an oxygen dissolution state, takes on a pink color in a slight oxygen dissolution state, and is colorless in an oxygen dissolution-free state; S2, adding L-cysteine hydrochloride, a biological buffering agent MES (morpholineethanesulfonic acid) and an anaerobic indicator into an anaerobic bottle with 1L of the fluid medium and mixing uniformly; S3, introducing nitrogen gas to the bottom of the anaerobic bottle in the S2 for 50-100min, pretreating a part of dissolved oxygen in the fluid medium to change the color of the fluid medium from the dark blue color to the light blue color or the light pink color; and S4, enabling the fluid medium of which a part of the dissolved oxygen is removed to stand, and consuming the rest of dissolved oxygen in the anaerobic bottle through the added L-cysteine hydrochloride in the S2 to change the color of the fluid medium from the light blue color or the light pink color to an achromatic color, thus completing deoxygenization. The deoxidizing method can be used for removing the dissolved oxygen in the fluid medium, and is conducive to enrichment and cultivation research of anaerobic micro-organisms.
Owner:ANHUI RUISIWEIER TECH

Method of screening 1-deoxyxylulose-5-phosphate racemase inhibitor from marine microorganism fermentation broth

The invention discloses a method of screening a 1-deoxyxylulose-5-phosphate racemase inhibitor from a marine microorganism fermentation broth. The method includes: 1) a step of preparing a marine microorganism fermentation broth sample, namely, a step of inoculating a fungus, fermenting, extracting metabolites, concentrating and eluting; 2) a step of designing a primer and performing PCR amplification by adopting the whole genome of vibrio vulnificus as a template, cloning a vector, converting escherichia coli, inducing expression of a target protein, and subjecting the target protein to elution, dialysis, ultrafiltration and desalting to obtain His-DXR; 3) a step of reacting a bacterial suspension, a resazurin solution and the marine microorganism fermentation broth sample obtained by the step 1), measuring the fluorescence value, calculating the inhibition rate through an equation, and screening the fraction the inhibition rate of which is higher than 80%; and 4) reacting the fraction screened in the step 3) and the His-DXR obtained in the step 2), calculating the inhibition rate, and screening the fraction the inhibition rate of which is not less than 50%. The method is simple and accurate, and lays a foundation for development of novel antibacterial medicines adopting Dxr as a therapeutic target or development of leading compounds.
Owner:MINJIANG UNIV

A method for screening 1-deoxyxylulose-5 phosphate racemase inhibitors from marine microbial fermentation broth

The invention discloses a method of screening a 1-deoxyxylulose-5-phosphate racemase inhibitor from a marine microorganism fermentation broth. The method includes: 1) a step of preparing a marine microorganism fermentation broth sample, namely, a step of inoculating a fungus, fermenting, extracting metabolites, concentrating and eluting; 2) a step of designing a primer and performing PCR amplification by adopting the whole genome of vibrio vulnificus as a template, cloning a vector, converting escherichia coli, inducing expression of a target protein, and subjecting the target protein to elution, dialysis, ultrafiltration and desalting to obtain His-DXR; 3) a step of reacting a bacterial suspension, a resazurin solution and the marine microorganism fermentation broth sample obtained by the step 1), measuring the fluorescence value, calculating the inhibition rate through an equation, and screening the fraction the inhibition rate of which is higher than 80%; and 4) reacting the fraction screened in the step 3) and the His-DXR obtained in the step 2), calculating the inhibition rate, and screening the fraction the inhibition rate of which is not less than 50%. The method is simple and accurate, and lays a foundation for development of novel antibacterial medicines adopting Dxr as a therapeutic target or development of leading compounds.
Owner:MINJIANG UNIV

High-sensitivity DNA fluorescence analysis method based on morpholine oligonucleotide functionalized magnetic microspheres

The invention discloses a high-sensitivity and high-selectivity fluorescent analysis method by introducing morpholine oligonucleotide to detect a specific DNA fragment. The method is as below: using morpholine oligonucleotide covalently attached to the surface of a magnetic microspheres as a capture probe, hybridizing the morpholine oligonucleotide with a long target DNA fragment; hybridizing an un-hybridized segment of the target DNA signal with a signal DNA with terminal modified with biotin; using thecombination effect of streptavidin and biotin, introducing alkaline phosphatase into the terminal of a signal probe DNA; catalyzing L-ascorbic acid 2-phosphate to generate L-ascorbic acid by alkaline phosphatase; and reducing resazurin by L-ascorbic acid to form strong fluorescent resorufin. The fluorescence intensity is positively correlated with the target DNA; through fluorescence analysis, high-sensitivity and high-selectivity detection of DNA fragments is achieved; and the method can be used in the detection of DNA samples in the fields of molecular recognition, clinical diagnosis, environmental monitoring and food safety.
Owner:NANJING UNIV OF SCI & TECH

Mycobacterium tuberculosis medicament sensitive phenotype detection method and application of method

The invention discloses a mycobacterium tuberculosis medicament sensitive phenotype detection method, which comprises the following steps of: 1) adding 2.5 to 6mul of 0.2 percent of resazurin dissolved by using methanol with mass percentage concentration into the inner side of a tube cover of a sterile centrifuge tube, and sterilizing the centrifuge tube after the methanol is volatilized; 2) inoculating 5 to 50mg of mycobacterium tuberculosis to be detected into a 7H9-S culture medium, regulating the concentration of the bacteria solution by adopting turbidimetry, regulating the concentration of the bacteria solution to the concentration of a Mcburney turbidimetric 1 tube by using sterile water or 0.9-percent physiological saline, and diluting the bacteria solution by 20 times by using the 7H9-S culture medium; 3) inoculating 50 to 150mu of diluted bacteria solution into the centrifuge tube of the step 1); 4) adding 100mul of anti-tubercular medicament solution diluted by 7H9-S and to be detected into the centrifuge tube of the step 3), wherein the concentration of the medicament to be detected is 0.01mug / ml to 2.5mg / ml; 5) after the centrifuge tube of the step 4) is covered closely, culturing the solution for 6 to 7 days at the temperature of 37 DEG C; and 6) inverting and oscillating the centrifuge tube of the step 5), and culturing the solution for 12 to 48 hours at the temperature of 37 DEG C after the solution becomes blue. Proved by experiments, in the mycobacterium tuberculosis medicament sensitive phenotype detection method and application of the mycobacterium tuberculosis medicament sensitive phenotype detection method in anti-tubercular medicament screening, the method has low workload and short detection time; compared with a traditional L-J medium medicament sensitive detection method, the accuracy reaches 90 to 99 percent; and the method has good biological safety, does not cause laboratory propagation of tuberculosis, is suitable for large-scale.
Owner:LANZHOU UNIVERSITY

Akermans myxobacteria culture medium and preparation method thereof

The invention discloses an Ackerman myxobacteria culture medium and a preparation method thereof. The Ackerman myxobacteria culture medium per liter is prepared from the following raw materials: 0.4g of monopotassium phosphate, 0.53g of disodium hydrogen phosphate, 0.3g of ammonium chloride, 0.3g of sodium chloride, 0.11g of calcium chloride, 0.1g of magnesium chloride hexahydrate, 4g of sodium bicarbonate, 0.5mg of resazurin, 1mL of an acid trace solution, 1mL of an alkali trace solution, 1mL of a vitamin solution, 37g of a brain heart infusion, 1.5g of xylooligosaccharide, 5g of cyclodextrin and 30g of solarbio protease-enzymolyzed egg white. The culture medium is capable of solving the problem of slow growth of Ackerman myxobacteria, and significantly improving the growth rate and the bacteria solution concentration of Ackerman myxobacteria.
Owner:INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI

Clostridium acetobutylicum strain and screening method and application thereof

The invention relates to a Clostridium acetobutylicum strain capable of quickly generating butanol by fermenting xylose and starch, and a screening method and application thereof. The invention discloses Clostridium acetobutylicum, which is classified and named Clostridium acetobutylicumXY16 with the preservation and registration number of CCTCC NO:M2010011. The strain obtained by ion beam mutation and screening of a xylose plate, a 2-deoxy-D-glucose plate, a bromocresol green plate and a resazurin plate can quickly and efficiently generate the butanol by fermenting xylose and starch, has the advantages of high yield of total solvent, high butanol ratio and good repeatability, and is a good strain suitable for industrial production.
Owner:NANJING TECH UNIV
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