145 results about "Ethidium bromide" patented technology

The invention discloses a preparation method of a marrow G-band for karyotype analysis in the medical examination field. The method includes adding ethidium bromide and demecolcine into a medium at the end of culture. The adding of a certain amount of ethidium bromide (EB) at the end of cell culture can make the chromosome in a split phase longer and the band patterns clearer. With the advantages of time saving, labor saving, and higher accuracy, the marrow G-band has more extensive application prospects in the field of medical examination.

Identifying primers and method of different groups of large yellow croaker, relates to a kind of primer and large yellow croaker. The invention provides an identifying primer (5'-TGCCGCACTT-3') for identifying 3 geographical species of Daiqu, west Min-yue, and Naozhou, and cultivation species. The identifying method comprises the steps of: extracting large yellow croaker DNA with phenol/chloroform, constructing a PCR system (25uL) containing Buffer 10mM, dNTP 0.2mM, Mg2+ 1.5mM, primer 0.2mM, Taq 1U,large yellow croaker DNA 50ng, and water as balance, detecting the product with agarose gel electrophoresis, dyeing with ethidium bromide, observing with UV, taking a picture, and recording. Different species are identified through comparing the result of electrophoresis and the provided standard graph.

The invention belongs to the field of molecular biology and discloses a pigeon sex discriminating method comprising the following steps of: designing an upstream primer SEQIDNO.1 (Sequence Identification Number 1) and a downstream primer SEQIDNO.2 by comparing a 2550F/2718R primer amplified product of a pigeon CHD gene with a CHD gene sequence landed on NCBI (National Center for Biotechnology Information); amplifying CHD gene fragments (preparing a genomic DNA template from micro feather pulp cells) on Z and W sex chromosomes of a pigeon by PCR (Polymerase Chain Reaction); dyeing by EB (ethidium bromide); judging sex according to a band type through 1 percent of agarose gel electrophoresis; appearing two bands, namely that 628bp and 358bp are male; and appearing three bands, namely that 628bp, 420bp and 358bp are female. The invention combines a new preparation method of an animal cell genomic DNA template and a high-efficiency pigeon CHD gene fragment amplimer, and the once detection rate and the detection accuracy of the pigeon sex discrimination can reach 100 percent.

The invention discloses a manufacturing method for marrow chromosome G band, and the method comprises adding mankind lymphoma cell cultures into a marrow medium and adding a certain amount of ethidium bromide when cell culture is terminated. The effects comprise that (1) division phases are relatively more, time is saved and labor is saved; (2) the length of chromosome in division phases is relatively long, and the band is relatively clear; (3) the dispersity is relatively good and the analysis is facilitated; and (4) the detection rate of abnormal chromosome is relatively high and the diagnosis result is relatively reliable. The G band manufactured by employing the method has the advantages of saving time and saving labor when applied to marrowchromosome karyotyping, is relatively high in accuracy and has relatively wide application prospect.

The invention discloses a detection primer and a detection kit for watermelon wilt disease fungi. The detection primer is a dual-detection primer set consisting of a wilt disease fungus universal primer and a primer which only can amplify a watermelon wilt disease fungi. A reagent of the detection kit comprises the detection primer, a buffer solution, dNTPs, Taq polymerase, watermelon wilt disease fungus positive control DNA and ultrapure water. The invention further discloses a detection method. The detection method comprises the following steps: (1) extracting DNA of a plant tissue infected by the watermelon wilt disease fungi; (2) carrying out PCR amplification on DNA which is separated in the step (1) by virtue of the detection kit; and (3) separating PCR amplification products in the step (2) by virtue of sepharose gel electrophoresis, dyeing the products with ethidium bromide under an ultraviolet lamp after the separation, and determining a result according to the sizes of the amplified products. According to the invention, a method for detecting watermelon wilt disease fungi at a biological molecular level is provided and has the characteristics of high precision and convenient operation.

The invention provides a polypropylene (PP) composition for automobile bumpers and a preparation method thereof. The PP composition for automobile bumpers is prepared from the following components in percentage by weight: 50-70% of PP copolymer, 10-20% of POE (polyolefin elastomer), 10-20% of talcum powder, 2-5% of PP-g-MAH (maleic anhydride), 1-2% of EB (ethidium bromide), 2-5% of flame retardant, 0.1-0.5% of photostabilizer and 0.1-0.5% of nucleating agent. The PP prepared according to the formula has the advantages of high low-temperature impact strength, high heat-deformation temperature, high toughness and ductility under high-speed stretch, low embrittlement tendency, high tensile strength, high tension strength, high shear strength, high flexural strength, high pressure intensity, low deformation, high stretching rate, high thermal expansion coefficient, low volatility, no migration, excellent ultraviolet resistance and excellent electric properties, and is suitable to be used as a material for automobile bumpers.

The invention relates to a loop-mediated isotheral amplification method (LAMP) for detecting Lyme disease pathogen in tick bodies, which is characterized by taking 16S rRNA gene as the target gene, using special software to design the LAMP for detecting the primers of the Lyme disease pathogen-Borrelia burgdorferi in the tick bodies, then selecting the primer of the specific fragment of the Lyme disease pathogen from the primers, extracting DNA after piercing the ticks to be detected, uniformly mixing the obtained DNA and the reaction buffer, the reaction mixture of the selected primer of the specific fragment and DNA polymerase to carry out amplification, adding loading buffer after activating the amplified product, then placing the mixture into agarose gel containing ethidium bromide to carry out electrophoresis detection and detecting whether the detected ticks carry the Lyme disease pathogen according to whether the specific band occurs after electrophoresis.

The invention relates to a molecular biological method for identifying gender of scandent hop, belonging to the technical field of the molecular biology. The specific implementing process is as follows: a designed specific primer sequence is utilized and a scandent hop individual genome DNA (deoxyribonucleic acid) is used as a template for PCR (Polymerase Chain Reaction) amplification, the reaction system is 25muL, the reaction compositions are as follows: 1U of TaqDNA polymerase, 2muL of dNTP (diethyl-nitrophenyl thiophosphate) and 100-200ng of genome DNA, the PCR amplification is performed on a PCR instrument, the electrophoretic separation is performed to the amplified product with 1.5% of sepharose gel, ethidium bromide is used for dying, the process is observed and recorded by photograph under a gel-imaging system, DL2000 marker is used as a molecular weight mark, the scandent hop with an objective stripe with size of 153bp is male, and the scandent hop without the stripe is female. The SCAR (sequence characterized amplified regions) primer is used for predicting the gender of the scandent hop, a simple and stable molecular marker is provided for identifying the gender of the scandent hop, and the molecular biological method has the important application value in the aspects of production and human health.

The utility model provides a method for effectively and quickly detecting high reproductive trait of sheep through BMP15 gene. The method comprises the following steps: 1) a SNP locus is newly found in a BMP15 gene coded sequence of Qira black sheep and positioned in a Genebank sequence, and a T-C mutation (T755C for short) occurs at a +755 locus by taking the initiator codon ATG of the BMP 15 gene sequence as the beginning; 2) a pair of nucleotide sequences are synthesized to serve as primers for detecting T755 C locus, the forward primer is 5'-gaagaccaaacctctccctaagg-3', the reverse primer is 5'-tactttcaggcccatcatgctcc-3', and both upstream and downstream primers are 23bp in length, the primers are combined on the corresponding template strands of the sheep BMP 15 gene, and amplified to the nucleotide sequence including an E+755 locus; 3) DNA is extracted, PCR is amplified through the primers, and the product is detected by 2% agarose gel electrophoresis and dyed by ethidium bromide, so that the amplified target fragment can be determined by gel imaging; and 4) the PCR product is digested by restriction enzyme Apal, and when the enzyme digestion product is fragments 120 bp and 140 bp, an individual is proven to have a high reproductive performance.

The invention provides a special prime designed according to conservative M genes of influenza virus. By the aid of the principle of specific binding of biotin and streptavidin, biotin-labeled PCR (polymerase chain reaction) products are bound on a microtiter plate, anti-digoxin peroxidase is added, and finally TMB (tetramethylbenzidine) color development solution is added, so that positive products can develop colors. On the basis, a special kit is successfully prepared. The method combines PCR with a high-sensitivity digoxin detection system, the sensitivity of the method is 100 times higher than that of a conventional agarose gel electrophoresis detection method, and the method is high in specificity and overcomes difficulty in detecting AIV (avian influenza virus) of a low content in a tissue sample. Compared with a virus isolation method, the method has the advantages that coincidence rate can reach more than 95%, pollution of chemical agents (such as ethidium bromide) is eliminated, automation is benefited, the method is suitable for detecting a large number of samples, and a new approach is provided for early diagnosis of avian influenza and molecular epidemiological investigation.

The invention belongs to the quick detection technology of food-borne pathogenic bacteria, and in particular relates to a detection method of loop-mediated isothermal amplification (LAMP) for salmonella. The detection method comprises the preparation of a reagent comprising a sample processing reagent, an LAMP reaction reagent, a large segment of BstDNA polymerase, agarose, bromophenol blue sampling buffer solution and ethidium bromide solution and the detection procedures of the detection an amplified product and the generation of deposition. The detection method comprises the following steps: firstly designing a specific primer; secondly extracting DNA from a sample; thirdly, establishing a reaction system. Compared with the traditional bacterial culture method, the immunity detection method and other molecular biology method adopted presently, the method has the advantages of simple operation, quickness, no special instrument reagent and specific sensitivity. The invention can be applied to the quick detection of the salmonella in foods and other samples.

The invention provides an SNP (Single Nucleotide Polymorphism) molecular marker for a sterile gene pms3 (photoperiod-sensitive male sterility 3) of paddy rice. The molecular marker is K-120522, wherein the polymorphism of the K-120522 is C or G. By utilizing the molecular marker, the gene pms3 can be quickly and precisely detected; the selection efficiency, in the breeding of the paddy rice, of the gene is greatly improved. The enzyme digestion, the electrophoresis, the sequencing and the like are not needed in a detection process; the operation is simple and convenient; the commercialized high-throughput quick detection can be realized; the aerosol pollution of a PCR (Polymerase Chain Reaction) product, the pollution of EB (Ethidium Bromide) to an environment and the harm of formaldehydeto a human body are thoroughly avoided.

The invention belongs to a rapid detection technology of food-borne pathogens, in particular to a loop-mediated isothermal amplification (LAMP) method for detecting shigella dysenteriae DNA. The method comprises the preparation of reagent comprising sample processing reagent, LAMP reactive reagent, a large fragment of Bst DNA polyase, agarose, bromophenol blue loading buffer solution and ethidium bromide solution and the procedures of the detection of amplification products and sediment generation. The method comprises the following steps: firstly, designing a specific primer; secondly, extracting sample DNA; and thirdly, preparing a reaction system. Compared with the currently adopted traditional bacterial culture method, an immunological detection method and other molecular biology methods, the method has the advantages of simple and rapid operation, no special instrument and reagent, sensitivity and specificity and can be applied to the rapid detection of shigella dysenteriae in foods and other samples.

The invention discloses a method for rapidly detecting live enterohemorrhagic Escherichia coli O157:H7. The detection method comprises the following steps: culturing Escherichia coli O157:H7 to obtaina bacterium suspension; adding an ethidium bromide monoazide solution into the bacterium suspension under a dark condition, standing in the dark, uncovering to place on ice, exposing, centrifuging, removing the supernatant, re-suspending with ultrapure water, centrifuging in a metal bath, and taking the supernatant as extracted bacterial genome DNA; designing and synthesizing primers and probes;diluting the DNA with sterile ultrapure water, and performing dual fluorescent PCR (Polymerase Chain Reaction) amplification; analyzing by selecting a 'maximum second derivative' method, and judging the results by the value CP. The method has the beneficial effects that the detection method is excellent in specificity and repeatability, high in sensitivity and accuracy, capable of rapidly identifying dead and live bacteria of the enterohemorrhagic Escherichia coli O157:H7 and low in detection cost.

The invention discloses a genotype inspection method and a genotyping inspection method of staphylococcal enterotoxin. The genotype inspection method of staphylococcal enterotoxin includes the steps of sample treatment, DNA extraction, PCR and PCR amplified product analysis. The amplified sample is put in agarose gel including ethidium bromide for electrophoresis and color development and observed under a transmissive uviol lamp; the inspected sample can be judged as a positive sample including corresponding staphylococcal enterotoxin genotype if the length of the amplified nucleic acid band is the same as the amplified length in table 2. The first method can fast inspect the particular staphylococcal enterotoxin genotype, with high sensitivity, high particularity and high accuracy. The second method can achieve systematic genotyping of staphylococcal enterotoxin genes from different sources and provide technical support for analysis of relationships among different enterotoxin types. The invention is significant in food security quarantine, disease diagnosis and molecular epidemiology research.

The invention relates to a gallinaceous visfatin gene 9bp indel polymorphism detection method and application thereof. The detection method comprises the following steps of: designing a primer in the position of a gallinaceous visfatin gene 9bp indel locus; then carrying out PCR amplification; and detecting the polymorphism of the gene 9bp indel by the polyacrylamide gel electrophoresis, wherein the gene is of a DD genotype when 9bp is deleted, the gene is of an II genotype when 9bp is inserted, and the heterozygous individual at the locus is of an ID genotype. The correlation analysis combining the detection results and economic characters of chickens is used for the assistant selection and the molecular breeding of chickens. The detection method does not need be subject to the enzyme digestion, thereby saving the cost and the time; the detection is carried out by adopting the polyacrylamide gel electrophoresis, thereby overcoming the limitation of the agarose electrophoresis to the small-fragment nucleotide difference detection. The method has the advantages of high resolution, high detection sensitivity and accurate genotype judgment; and the gel has on requirements for the staining method, and the ethidium bromide (EB) staining and the silver staining are both applicable.

The invention discloses a detection kit and a detection method for viable bacteria in vibrio parahaemolyticus in food. The kit comprises 10* reaction buffer, a Bst DNA polymerase, dNTPs, MgSO4, DNase/RNase-FreeddH2O, a primer F3, a primer B3, a primer FIP, a primer BIP, a primer Loop-F, a primer Loop-R, ethidium bromide monoazide and 100*SYBRGreenI fluorescent dyes. The detection method includes steps of inoculating animality aquatic products animals and products to sodium chloride alkaline peptone water for overnight suspension culture after preprocess of samples; adding the ethidium bromidemonoazide to overnight suspension culture, placing the overnight suspension culture in a dark place, and performing the whole photoexcitation process on ice; and extracting sample DNA by using a DNA rapid extracting method, namely boiling bacteria suspension processed by the ethidium bromide monoazide, performing centrifugation, taking supernatant after the centrifugation as a DNA template of a loop-mediated isothermal amplification reaction, and performing the loop-mediated isothermal amplification reaction. The detection kit and the detection method for viable bacteria in vibrio parahaemolyticus in food have the advantages of being rapid, efficient, high in accuracy and sensitivity, convenient to use on sites, and widely suitable for detection for food, sanitation, immigration and the like.

The invention relates to a method for in-situ detecting Hg<2+> in a water sample through a photocatalysis visible manner. The method is characterized in that Hg<2+> is specifically combined with thymine (T) to form a -Hg<2+>-T structure, so that two T-enriched CpG-oligodeoxynucleotides are matched with each other in a staggering manner to form double strands. ethidium bromide (EB) can be embedded in the double strands, and 3,3',5,5'-tetramethyl benzidine (TMB) is oxidized under the radiation of a green LED (light emitting diode) lamp into blue. The semi-quantitative analysis of mercury ions can be realized by observing the color variation of the solution. By adopting the method, 0.05ngmL<-1> of Hg<2+> in the water sample can be detected through eyes, and the determination requirement of the Hg<2+> (1ngmL<-1>) in water samples such as drinking water, irrigation water for farmland and the like can be met. The method is good in selectivity, high in sensitivity, capable of avoiding the complicated separation and enriching process and directly applicable to the in-situ analysis of Hg<2+>.

The invention discloses a method for rapidly identifying opogona sacchari (an entry plant quarantine pest), which comprises: performing PCR amplification of the extract of the genome DNA of a sample to be tested by using opogona sacchari specific primer CF720/CR1113; performing electrophoresis on PCR amplification solution in 0.5XTE electrophoresis buffer under a voltage of 5V per centimeter of gel; and placing the PCR amplification solution in an ethidium bromide staining box for staining, and observing an amplification band with an ultraviolet gel imaging device to determine if the tested is opogona sacchari (the entry plant quarantine pest). The method has the advantages that: the identification time is reduced to less than 24 hours from the original more than one week; and the identification accuracy is improved to 99 percent from the original 70 percent. The method can help entry and exit quarantine and inspection department, customs and other related units to perform quick, accurate and high efficiency detection and identification at ports.

The invention discloses a full-automatic glue-preparation electrophoresis ultraviolet glue sweeping and cutting instrument which comprises a box body. The box body is divided into three layers, a baffle is arranged in the box body, the baffle divides the interior of the box body into six cells on the left side and the right side, the upper part of the box body is provided with a panel, and a liquid crystal screen, a numerical control panel and a control circuit respectively connected with the liquid crystal screen and the numerical control panel are arranged on the panel of the box body respectively; the left side and the right side of the second layer of the box body are respectively provided with a periscope view port; and in the six cells, an elevator, a drying device, an automatic cleaning device, an air-exhaust liquid-exhaust device, a mechanical arm, a plastic gel supporting plate, an adjustable glue preparation tank, a stirring rod, microcontrol integrated automatic glue preparation equipment, a miniature sample adding gun, an ethidium bromide container, a liquid adding device, a periscope, an electrophoresis tank, an ultraviolet glue sweeping camera, a glue cutting instrument, a centrifuge tube clamp and an ethidium bromide dye vat are comprised. The full-automatic glue-preparation electrophoresis ultraviolet glue sweeping and cutting instrument disclosed in the invention has the advantages of automation, integration, convenience and quickness, and greatly increases the efficiency, the precision and the safety of experiments.

The invention relates to the method used PCR to identify the goat early embryo sex. It includes the following steps: sampling from goat early embryo sex to make template DNA; designing primer according to goat SRY gene order; adding the template DNA, primer, DNA polymerase, reaction buffer solution, and dNTP into reaction tube to use as reactant; repeating denaturalization-annealing-stretching three steps to the end of the reaction. Its features are that it adds ethidium bromide into the reaction tube before PCR, directly observes the result after PCR by putting the reaction tube under ultraviolet lamp. It designs a pair of primers according to goat Y chromosome specificity sequence, identifies by non-electrophoresis PCR, simplifies operational programs, and is convenient for popularization and application in production.

The invention discloses a method for rapidly detecting virus of a potato tuber. The method is characterized by comprising the following steps of: extracting the RNA of the potato tuber, performing reverse transcription-polymerase chain reaction (RT-PCR), detecting PCR products through a nucleic acid colloidal gold chromatography test strip, and the like. The method suitable for rapidly detecting the virus of the potato tuber is established by combining the RT-PCR technology with the nucleic acid colloidal gold chromatography technology on the premise of optimizing a method for extracting the RNA of the virus of the potato tuber. The method has the greatest characteristic that: an RNA extracting solution is developed, the RT-PCR detection can be directly performed after a potato sample to be detected is ground, an RNA extraction step in the conventional method is eliminated, and the problem that the RNA is difficult to extract due to high starch content of the potato tuber is solved; and moreover, the developed nucleic acid colloidal gold chromatography test strip can finish the detection of the PCR products within ten minutes, electrophoresis and the dyeing of ethidium bromide are not required, and the method is rapid, practical, safe and pollution-free.

The invention discloses a method for detecting ethidium bromide by a fluorescent carbon dot probe. The method is characterized in that a fluorescent carbon dot is used as the probe, so that high-sensitivity detection is carried out based on the characteristic that fluorescence intensity of the fluorescent carbon dot is weakened when the concentration of ethidium bromide solution is increased, the change value of the fluorescence intensity of the fluorescent carbon dot and the ethidium bromide concentration form a good linear relationship, and the relevant coefficient is 0.999. The method is simple in operation, fast in detection, high in sensitivity and excellent in selectivity, thus rapid online in-situ sensitive detection can be executed to ethidium bromide in a mixed sample by the method.