31 results about "GeneQuant" patented technology

The invention provides a method for detecting Escherichia coli in milk based on an ultraviolet-visible spectrum technology. Under the complex background environment similar to that of the milk, the problems that detection spectra of microorganisms are affected by unfavorable factors, the phenomena of characteristic wavelength red shift, obvious noise and the like are generated, consequently spectral characteristics are difficult to pick up, and the accuracy of quantitative detection results is low can be effectively solved. The method comprises the steps that firstly, Escherichia coli powder is activated and subcultured, and a to-be-detected bacterium suspension is prepared; an ultraviolet spectrophotometer obtains ultraviolet-visible spectra of the to-be-detected bacterium suspension at different culture times; noise interference is eliminated through a standard normal correction method and an S-G convolution smoothing method; the spectral characteristic red shift phenomenon is explained from the angle of a formation mechanism of a benzene ring conjugated structure, and thus the characteristic waveband range is determined; the characteristic wavelengths are picked up in the characteristic waveband through a continuous projection algorithm, and the characteristic wavelengths are effectively screened according to the secondary growth law of the microorganisms; and the model relationship between the characteristic wavelengths and the total number of the microorganisms is established through a partial least squares method, and the total number of the Escherichia coli in the milk is quantitatively analyzed

The invention discloses a reovirus-detecting fluorescence quantitative PCR kit and an application thereof. The kit comprises: a) an RNA extraction solution, b) a reverse transcriptase reaction solution, c) reverse transcriptase, d) an RNA enzyme inhibitor, e) a primer and a TaqMan probe, f) a standard positive DNA template and g) a PCR fluorescence quantitative reaction solution. The kit is characterized in that: primer sequence is a sense primer: 5'-TGCGCCTATCCTTGAGTTGA-3', and an antisense primer: 5'-TTGCCAGGAAATACGGGTCT-3', and the size of an amplicon is 138 bp; the sequence of a fluorescence probe is: 5'-FAM-TCAAAATGGTGGACTTCAGTTTCGATTT-TAMRA-3', the 5' end of the probe is marked with a fluorescence emission group FAM, the 3' end is marked with a fluorescence quenching group TAMRA, the standard positive DNA template transforms a colon bacillus DH5 alpha by a pGEM-T carrier inserted with reovirus S1 protein zone 361 bp segment, plasmid is extracted after proliferation, an A260 ration is measured in an ultraviolet spectrophotometer, and the plasmid is diluted by 10 times of gradient. The kit can efficiently and conveniently monitor the reovirus pollution in serum products in real time, can be applicable to epidemiology investigation of reovirus infection, and can provide technical support for relevant fundamental researches, thus having quite broad application prospect.

The invention discloses a fluorescence quantitative PCR kit for detecting calf diarrhea virus and application. The kit comprises a) RNA extract, b) reverse transcriptase reaction liquid, c) reverse transcriptase, d) RNA enzyme inhibitor, e) primers and TaqMan probe, f) standard positive DNA template, and g) fluorescence quantitative PCR reaction liquid. The kit is characterized in that in a positive primer, a negative primer and a fluorescence probe sequence, a 5' end of the probe marks a fluorescence emitting group FAM, a 3' end of the probe marks a fluorescence quenching group TAMRA, the standard positive DNA template converts colon bacillus DH5a by a pGEM-T carrier inserted into a 185bp fragment of a calf diarrhea virus 5'-UTR area, plasmids are extracted after multiplication, and A260 is measured by an ultraviolet spectrophotometer to definite quantity and is diluted by 10 times of gradient. Preparation for the kit comprises the following steps: A, treatment of a specimen and a standard product; and B, RT-PCR amplification by a two-step method and real-time fluorescence detection, and application of the fluorescence quantitative PCR kit in medicaments for quantitatively detecting calf diarrhea virus. The quantitative result is more accurate, reliable and stable, and the repeatability is good.

The invention discloses a bovine parainfluenza type-3 virus detecting fluorescence quantitative PCR kit and an application thereof. The kit comprises: a) an RNA extraction solution, b) a reverse transcriptase reaction solution, c) reverse transcriptase, d) an RNA enzyme inhibitor, e) a primer and a TaqMan probe, f) a standard positive DNA template, and g) a fluorescence quantitative PCR reaction solution. The kit is characterized in that primer sequence is respectively a sense primer and an antisense primer: the size of an amplicon is 129 bp; in the sequence of a fluorescence probe, the 5' end of the probe is marked with a fluorescence emission group FAM while the 3' end is marked with a fluorescence quenching group TAMRA; the standard positive DNA template transforms a colon bacillus DH5 alpha by a pGEM-T carrier inserted with bovine parainfluenza type-3 virus conservative F protein coding zone 188 bp segment, plasmid is extracted after proliferation, an A260 ration is measured in an ultraviolet spectrophotometer, and the plasmid is diluted by 10 times of gradient. The result of the kit is more accurate, reliable, stable and repeatable. The kit can also be applied to the epidemiology investigation of bovine epidemic influenza and provide technical support for relevant fundamental researches, thus having quite broad application prospect.

The invention discloses a rabies virus detecting fluorescence quantitative PCR kit and an application thereof. The kit comprises: a) an RNA extraction solution, b) a reverse transcriptase reaction solution, c) reverse transcriptase, d) an RNA enzyme inhibitor, e) a primer and a TaqMan probe, f) a standard positive DNA template, and g) a PCR fluorescence quantitative reaction solution. The kit is characterized in that: primer sequence is a sense primer: 5'-TAGGATGCTATATGGGTCAAGTCAGA-3', and an antisense primer: 5'-TTCAAATGTCCCTTTCCCGAAGAA-3', and the size of an amplicon is 125 bp; the sequence of a fluorescence probe is: 5'-FAM-CAACGGTTATTGCTGCATGTGCTCCTGA-TAMRA-3', the 5' end of the probe is marked with a fluorescence emission group FAM, the 3' end is marked with a fluorescence quenching group TAMRA, the standard positive DNA template transforms a colon bacillus DH5 alpha by a pGEM-T carrier inserted with rabies virus N protein zone 391 bp segment, plasmid is extracted after proliferation, an A260 ration is measured in an ultraviolet spectrophotometer, and the plasmid is diluted by 10 times of gradient. The kit can efficiently and conveniently monitor the rabies virus pollution in serum products in real time, and can provide technical support for relevant fundamental researches, thus having quite broad application prospect.

The invention provides a method for detecting zymoprotein residues in cefprozil prepared by an enzymatic method. The method is characterized by comprising the following steps: preparing a Coomassie brilliant blue G-250 dye solution and a standard protein stock solution; preparing a standard curve solution by taking a sodium bicarbonate solution as a solvent; preparing a blank solution by taking asodium bicarbonate solution as a solvent; preparing a test sample determination solution by taking a sodium bicarbonate solution as a solvent; drawing an absorbance-protein concentration standard curve through blank solution correction by using an ultraviolet spectrophotometer; and calculating the residual content of zymoprotein contained in the test sample according to the absorbance value read by the test sample determination solution. Sodium bicarbonate is ingeniously selected as a solvent, cefprozil can be dissolved, zymoprotein can be fully dissolved, the residual quantity of zymoproteinin a cefprozil product prepared through an enzymatic method can be effectively measured, the medication safety of medicine is improved, and the detection method is easy to operate, high in accuracy and good in reproducibility and reliability.

The invention relates to a near-infrared fluorescent probe for detecting selenocysteine (Sec), and concretely relates to an organic compound based on cyanine and an application thereof. A structural formula of the organic compound based on the cyanine is shown as a formula I, the structural formula of the compound is shown as the formula I in the specification, and the compound is taken as a fluorescent probe of Sec. The Sec fluorescent probe compound can generate obvious displacement of corresponding fluorescence wavelength under existence of Sec, which can be used for Sec detection, interference by external detection condition is greatly reduced, and detection precision is increased. Under existence of Sec, according to the Sec fluorescent probe compound, the ultraviolet absorption generates obvious change, and an ultraviolet spectrophotometer and naked eyes are simultaneously used for detection. The compound as the fluorescence probe can be used for detecting the Sec level inside and outside cells, and has important biomedical meaning on a deep research a dynamics mechanism of generation, conveying and accumulation of Sec in organism, especially the research of physiological effect of Sec in mitochondria.

The invention discloses a T3 DNA ligase-based cadmium ion detection method, which comprises the following steps: designing a DNA template, and when a target object cadmium ion exists, reducing the distance between two ends of a chain by the target object, so that a 3'end and a 5 'end are complementarily paired to form a blunt end; under the action of T3 DNA ligase, cyclizing the template; adding a section of oligonucleotide chain complementarily paired with the annular template as a primer for subsequent amplification reaction; under the action of adding phi 29 DNA polymerase, starting a rolling circle amplification reaction to obtain a longer nucleotide chain; adding a fluorescent dye SYBR Green I into the obtained product, inserting the dye into a DNA chain, detecting the sample by using an ultraviolet spectrophotometer under the excitation wavelength of 517-537 nm, and then measuring the fluorescence intensity under the excitation wave of 487-507 nm; detecting the cadmium ions by using a Real-Time PCR method, and the accuracy of the method disclosed by the invention is verified.

The invention provides an enzyme-assisted hairpin probe remodeled label-free colorimetric sensor based on target trigger and an application of the sensor. The invention also provides a method for visual detection of nucleic acid. According to the method, the nucleic acid is used as a target, a reaction is performed with the help of enzymes, G-quadruplex sequences that can produce colorimetric signals are formed through multiple ways, and the G-quadruplex sequences can be used for visual detection of the nucleic acid; the method controls the reaction conditions, a hairpin probe (HGP) opened bya target gene under the action of a T4 DNA ligase is connected with another double-stranded DNA (DGP) to complete remodeling, under the synergy of a DNA polymerase and a restriction enzyme, the targetDNA is recycled, and multiple channels for generating the G-quadruplex sequences are generated. According to the method, the enzymes are used to assist the DNA reaction to form a large amount of G-quadruplexes, and the G-quadruplexes can be used for detection by using an ultraviolet spectrophotometer, and can be used for visual detection by using naked eyes; and the detection method is convenient, the materials used in the method are commercialized, and the biological safety is high.

The invention discloses a pretreatment method for determination of 18 amino acids in C57 mouse brain tissue and plasma and belongs to the field of pretreatment of endogenous substances in biological samples. According to the method, an ultraviolet spectrophotometer and a high performance liquid chromatograph are used, a method for treating brain tissue homogenate and plasma is investigated by combining liquid-liquid extraction with a protein precipitation method, and the method mainly investigates the influence of a sample pretreatment method on the removal amount of proteins, the removal amount of total lipids and the extraction amount of 18 amino acids in a sample, and further optimizes to obtain the optimal pretreatment method for determining 18 amino acids in C57 rat brain tissues andplasma.

The invention discloses a method for constructing a perilla frutescens core germplasm resource library based on an SRAP molecular marker, and belongs to the technical field of genetic diversity evaluation of perilla frutescens germplasm resources and construction of core germplasm. The method comprises the following steps of screening out 100 parts of perilla frutescens variety materials, planting the perilla frutescens variety materials in a flowerpot, taking seedling plant leaves as test materials, extracting total DNA of each variety, detecting the purity and integrity of the total DNA of each variety by using 1% agarose gel electrophoresis, determining the light absorption values of the total DNA of each variety at 260nm and 280nm by using an ultraviolet spectrophotometer, determining the purity and concentration of the DNA, and storing at -20 DEG C for later use; and carrying out SRAP-PCR amplification by using the total DNA, and carrying out electrophoretic separation on the amplification product by using 6% polyacrylamide gel. According to the method, a fingerprint spectrum is constructed for 100 parts of perilla frutescens germplasm resources by utilizing an SRAP marking technology, a perilla frutescens molecular retrieval table is compiled, core germplasm construction is carried out, the genetic relationship between perilla frutescens species and cultivated varieties is disclosed, a scientific classification system is established, and genetic diversity evaluation is carried out on the perilla frutescens perilla species and cultivated varieties.

The invention discloses a method for detecting hot spot mutation of a human TERT gene promoter. The method comprises the following steps: a, enriching templates by adopting DNA enrichment conventionalPCR aiming at different specimens; b, purifying the PCR product, determining the concentration by using an ultraviolet spectrophotometer, and performing concentration calibration and standard substance preparation; and c, carrying out TERT-146 (C/T) and TERT-124 (C/T) mutation rate quantitative detection on the calibrated specimens by adopting qPCR. According to the method, a qPCR technology platform which is conventionally developed clinically is adopted, the high-sensitivity TERT hot spot mutation rate quantitative detection method is established for DNA specimens from various sources, thelower limit of the detected mutation rate reaches 0.05%, and the method has the advantages of being high in detection sensitivity, high in specificity, capable of accurately and quantitatively detecting the mutation rate, low in cost, suitable for conventional clinical development and the like.

The invention relates to the technical field of polypeptide decoloration, in particular to a method for measuring the decoloration effect of a protein polypeptide solution. The method comprises the following steps: (1) dissolving dry polypeptide samples before and after decoloration in ultrapure water or deionized water according to the same concentration ratio to obtain a sample solution, standing at room temperature for a certain time, respectively scanning the sample solution and the ultrapure water at 380nm-780nm at an interval of 1-10nm by adopting a microplate reader or an ultraviolet spectrophotometer, and measuring the absorbance of the sample solution and the ultrapure water in the range; (2) making a wavelength-absorbance broken line graph; and (3) determining the decolorization effect according to the change trend of the broken line graph or the calculated decolorization rate through the change of qualitative or quantitative indexes. The method for measuring the decolorization effect of the protein polypeptide solution makes up the defects in the research in the prior art, the decolorization effect is observed more visually, and the method is suitable for solutions which are difficult to distinguish visually and have small differences.

The invention relates to the field of chemical measurement, in particular to a method and system for measuring the anti-Xa factor titer of a low-molecular-weight heparin calcium injection.The method comprises the steps that 100 microliters of a low-molecular-weight heparin calcium solution and 100 microliters of an antithrombin solution are taken and evenly mixed, balancing is conducted for 1 minute at the temperature of 37 DEG C, and a first mixed solution is obtained; adding 200 [mu] l of an FXa solution into the first mixed solution, uniformly mixing, and balancing at 37 DEG C for 1 minute to obtain a second mixed solution; adding 500 [mu] l of a chromophoric substrate solution into the second mixed solution, mixing, and balancing at 37 DEG C for 4 min to obtain a test solution; continuously measuring the absorbance of the test solution at the wavelength of 405nm by using an ultraviolet spectrophotometer and taking water as a blank, recording the current absorbance every one minute, totally 5 minutes, and calculating the absorbance change rate; 100 [mu] l of a trihydroxymethyl aminomethane-sodium chloride buffer solution and 100 [mu] l of a test solution are taken to replace a low-molecular-weight heparin calcium solution to be operated according to the steps, the absorbance change rates are calculated respectively, operation is more convenient, and the working efficiency is improved.

The invention relates to a hydrogen sulfide colorimetric sensor for targeted induction of mimic enzyme inactivation based on a mixed node metal organic framework material, and belongs to the technical field of biosensor environment detection. The synthetic Cu-Fe MOFs with mixed nodes are used as an artificial mimic enzyme to catalyze H2O2 to oxidize 3,3,5,5-tetramethylbenzidine (TMB) to generate oxTMB, and the solution is changed from colorless to blue; when dissolved H2S is added, H2S and Cu 2+ have strong binding capacity under an acidic condition; therefore, the mimic enzyme is inactivated, so that the capability of oxidizing TMB by H2O2 is weakened. then, along with the increase of the H2S concentration, the catalytic activity of the mimic enzyme is gradually reduced, and the color is obviously and gradually lightened, so that the qualitative or quantitative detection of H2S can be realized through an ultraviolet spectrophotometer or the color change. Based on the concerted catalysis of Cu-Fe in the Cu-Fe MOFs, the sensitivity and selectivity of H2S determination are improved. The method has a good application prospect in water quality analysis and environmental protection.