253 results about "DNA extraction" patented technology

The invention discloses an extraction preparation method and application of algae micromolecule PDRN, and relates to the technical field of biology. The invention establishes a process for extracting PDRN from microalgae, which comprises the following steps: carrying out cell disruption on microalgae liquid, and removing cell debris to obtain filtrate; the filtrate is subjected to DNA dissolution, DNA separation and DNA purification, and a PDRN solution is obtained; wherein the dry weight of the microalgae in the algae liquid is 0.1 to 0.333 g / mL. Through process optimization, the extraction efficiency and purity of the microalgae PDRN are improved, and the extracted PDRN has excellent anti-aging and anti-inflammatory effects.

The invention discloses a method for evaluating enhancement and release seedling resources based on environmental DNA polymerization analysis. The method comprises the following steps: carrying out gridding partition on a target water area, collecting a water sample through a designed sampling scheme, and carrying out DNA extraction and high-throughput sequencing to obtain species sequence information of each sampling point. Sequencing data is subjected to species identification by using a bioinformatics method, released species are identified, a spatial abundance model is established, and a preliminary distribution map is generated. And establishing a DNA degradation kinetic model in combination with water area environmental parameters, and carrying out reverse correction on abundance distribution. And through a resource inversion model coupled with hydrodynamics, analyzing biomass distribution characteristics and migration laws of the release group, and obtaining a resource evaluation result. And finally, a species environment preference model is constructed based on migration path analysis, an optimal release area is matched in a target water area, a scientific scheme including release point locations, opportunities and quantity is generated, and a whole-process technical support and a decision basis are provided for enhancement and release.

The invention provides a method for deducing the abundance of each family in a group based on mixed pool simplified genome sequencing, which comprises the following steps of: (1) establishing a parent 2b-rad library, and extracting SNP (Single Nucleotide Polymorphism) genotypes and sites by using a 2b-rad standard process; combining the SNP genotypes of the 2n parents into genotypes of n parent pairs according to a male parent and female parent relationship; comparing the genotypes of the parent pairs, and determining family specific tags; (2) extracting offspring individual tissues, mixing, and carrying out DNA extraction and 2b-rad library building sequencing on the mixed sample tissues; counting the sequencing depth of all the specific tag sites and calculating the secondary allele frequency of the specific tag of the mixed sample filial generation; and (3) calculating the proportion of each family in the offspring according to the sequencing depth of the specific tag site and the secondary allele frequency of the specific tag of the mixed offspring. According to the method, the overall feature distribution of the family can be accurately evaluated without sequencing each individual, so that the cost is greatly reduced.

The invention relates to the technical field of crop molecular breeding and detection, in particular to a sweet waxy corn molecular detection system based on an sh1 gene InDel marker and application of the sweet waxy corn molecular detection system, the sweet waxy corn molecular detection system is composed of a forward primer, a reverse primer and a transposon specific primer, and the sequences are shown as SEQ ID NO: 1-3. The sh1 gene InDel marker-based sweet waxy corn molecular detection system provided by the invention can be used for corn sh1 gene Mu insertion rapid detection and / or corn molecular marker-assisted sweet waxy corn breeding. By adopting a sweet waxy corn molecular detection system based on the sh1 gene InDel marker, the accuracy rate is 98.7%. And the detection cost of each sample is 0.15 dollars, and compared with the traditional SSR marking technology, the cost is reduced to 3% of that of the traditional SSR marking technology. From DNA extraction to final result interpretation, the whole detection process takes 3 hours.

The present invention discloses a microecological animal model for obstructive sleep apnea syndrome, which includes steps such as constructing a fitting model for the changing trend of OSA events, selecting target animals and performing pretreatment, preparing an animal model, obtaining fecal samples, and DNA extraction and 16S rRNA high-throughput sequencing. By constructing a fitting model for the changing trend of respiratory sleep apnea events, the present invention obtains the fitting mathematical relationship between the number of apnea and time, and establishes an animal model with SD rats. According to the obtained fitting mathematical relationship, the animal model of the experimental group is given periodic oxygen supply to establish an intestinal flora model for studying obstructive sleep apnea syndrome, which can simulate the actual physiological environment during sleep of patients. According to the changing trend of the duration of patients' respiratory sleep apnea events, a microecological animal model more consistent with the actual situation is established, making the detection result of the flora structure more accurate.

The invention provides a wheat gene sample automatic extraction device, which comprises an extraction processing box, supporting foot bases, a sealing door, an automatic control panel group, a vibration crushing box, a cooling sealing cover, an insulating handle, a cooling liquid nitrogen tank and an analysis detection box, the supporting foot bases are respectively in bolted connection with four corners of the lower part of the extraction processing box; the sealing door hinge is connected to the lower side of the front part of the extraction processing box; the automatic control panel set is connected to the upper side of the front portion of the extraction processing box through screws. Liquid nitrogen grinding is combined with ceramic grinding beads, and a vibration motor, a supporting tray, a locking screw, a test tube clamping seat and a constraint clamping hole are utilized, so that high-throughput treatment of multiple wheat samples can be realized at the same time, the DNA extraction cost of a single sample is reduced by 60%, and cells are uniformly broken; and the analysis detection box, the detection probe group, the cracking bin, the communicating pipe and the processor are arranged, so that different tissues such as wheat endosperm (high starch) and leaves (high fiber) can be processed.

The invention discloses a pretreatment method for panda excrement DNA extraction and application, and relates to the technical field of molecular biology. The pretreatment method comprises the following steps: loading newly collected excrement into a sterile sampling bag, so that a sample is quickly and completely frozen to be hard, then breaking the excrement into pieces, freeze-drying, sieving, sealing and cryopreserving. The pretreatment method can enrich DNA of a surface mucous membrane layer and trace DNA in excrement, does not need to peel off the surface mucous membrane layer, and is time-saving and labor-saving. The pretreatment operation is simple, and no reagent is needed. The pretreated excrement is small in size, good in stability, easy to store and capable of being stored in a refrigerator for a long time for standby application, and the needed storage space is small. The effective storage life and the use period of the sample can be prolonged, and technical support can be provided for molecular and genetic research of pandas.

The invention discloses a mountainous area surface matrix physical property prediction method, and relates to the technical field of geological exploration, and the method comprises the steps: obtaining mountainous area surface point cloud data, separating vegetation from ground points through a point cloud classification algorithm, and generating a digital elevation model and a digital surface model; based on the digital elevation model and the digital surface model, a quantum gravity gradiometer is adopted to move along a contour line for measurement, a gravity gradient tensor is obtained, terrain interference signals are eliminated, and a vertical density profile is obtained through inversion; identifying a density abnormal region according to the vertical density profile, acquiring an undisturbed soil column sample by adopting a drilling unit, determining a grain composition and moisture characteristic curve, performing DNA extraction and microorganism sequencing on the undisturbed soil column sample, and calculating the relative abundance of the nitro spirillum phylum. According to the method, the dynamic coupling model is constructed through fusion of quantum gravity gradient measurement and microbiological omics data, and the problem of insufficient vertical density-permeability coefficient coupling modeling precision of a traditional method is solved.

The invention relates to a method for extracting genome DNA of plants rich in secondary metabolites and a buffer solution, and belongs to the technical field of molecular biology and botany. The method solves the technical problems of low DNA extraction efficiency, poor purity, easy degradation and the like when a traditional DNA extraction method is used for treating plant tissues rich in secondary metabolites such as alkaloid, polyphenol and the like. Comprising the following steps: adding a complexing agent such as polyethylene glycol or polyvinylpyrrolidone when grinding plant tissues in a liquid nitrogen environment; splitting by using a cell wall splitting buffer solution containing dithiothreitol and a nonionic surfactant; carrying out DNA release and extraction at 60-70 DEG C by adopting a CTAB (Cetyltrimethyl Ammonium Bromide) extraction buffer solution containing polyethylene glycol and papain; and then purifying and precipitating to obtain high-purity genome DNA (Deoxyribose Nucleic Acid). The method can effectively remove alkaloid, protein and other impurities, significantly improves the DNA yield and purity, and is suitable for genome sequencing, genetic resource protection, medicinal plant molecular identification and the like of plants with high secondary metabolites such as Stephania kwangsiensis and the like.

The invention relates to the technical field of biology, and particularly discloses a gene Panel sequencing method for intrahepatic cholestasis in a gestation period, which comprises the following steps: collecting saliva, performing DNA extraction and purification on a sample, breaking the purified sample, treating the broken sample to obtain a supernatant A, performing linker connection and purification on the supernatant A to obtain a supernatant B, and sequencing the supernatant B to obtain the gene Panel of intrahepatic cholestasis in a gestation period. The invention relates to a Panel detection technology for detecting 186 cholestasis genes covering bile acid metabolism, which comprises the following steps: preparing a supernatant B, carrying out pre-PCR and purification treatment on the supernatant B to obtain a pre-PCR product, carrying out liquid phase hybridization and capture, configuring a PCR reaction system, carrying out Post-PCR reaction and purification to obtain a supernatant C, quantifying the supernatant C, screening ICP susceptible genes through mixed sample sequencing, collecting DNA (Deoxyribonucleic Acid) through a saliva sampling mode, and detecting the 186 cholestasis genes covering bile acid metabolism. Other hereditary intrahepatic cholestasis diseases can be excluded, and ICP genetics susceptibility genes are defined.

The invention provides a lysis binding solution, an extraction kit and a detection method for extracting residual DNA of host cells, the lysis binding solution comprises guanidine hydrochloride, PEG6000, tRNA, glycogen and isopropanol, the concentration of guanidine hydrochloride is 2.0-2.5 M, the concentration of PEG6000 is 1%-10% (m / v), and the concentration of isopropanol is 50-60%. According to the present invention, the host cell residual DNA extraction is performed by using the lysis binding liquid, such that the compatibility between the reagent and different sample types can be significantly enhanced, the sample pre-dilution is reduced, and the matrix interference is reduced so as to improve the extraction efficiency and the extraction accuracy of the residual DNA, and effectively overcome the limitation of the commercial kit.

A sampling device including an intake pump coupled with a volume controller to provide a known volume of samples such as liquid collected through a specialized hollow membrane filter. Once the known volume of sample is filtered the intake pump is turned off, an air pump turns on and removes residual or excess sample. The membrane filter is then preserved for extraction of DNA / RNA.

The present application relates to a deoxyribonuclease mutant and its application in preparing PDRN. The amino acid sequence of the deoxyribonuclease mutant is shown as SEQ ID NO. 1, and the nucleotide sequence of the encoding gene is shown as SEQ ID NO. 2. The present application also provides the application of the deoxyribonuclease mutant and / or the encoding gene of the deoxyribonuclease mutant in preparing PDRN, and the specific application method. The present application is based on the deoxyribonuclease expressed by the DNASE1 gene of bovine pancreas tissue, and the amino acids at positions 69, 132 and 215 are selected for iterative saturation mutation. Compared with the wild-type enzyme, the enzyme activity of the deoxyribonuclease mutant A69H-S132R-Q215C is increased by 2.15 times, reaching 4.1x10 6 U / mL, and the DNA extraction rate of salmon PDRN prepared by using the mutant enzyme prepared by the present application is 15.2%, the DNA content is 99.1%, and the molecular weight is 1.05 million Daltons, which is significantly improved compared with the wild-type enzyme, and the PDRN in this molecular weight range has more significant biological activity.

The invention discloses a mitochondrial molecular marker for identifying acrossocheilus fasciatus from different water systems and application of the mitochondrial molecular marker. A primer kit of the acrossocheilus fasciatus mitochondrial molecular marker comprises a primer pair as shown in SEQ NO: 1 and SEQ NO: 2, and / or a primer pair as shown in SEQ NO: 3 and SEQ NO: 4; the acrossocheilus fasciatus mitochondrial molecular marker is formed by amplifying primers contained in the acrossocheilus fasciatus mitochondrial molecular marker primer kit. The identification method comprises the following steps: carrying out DNA extraction on acrossocheilus fasciatus to be identified; carrying out PCR (Polymerase Chain Reaction) amplification on the extracted DNA target fragment by using the primer pair; sequencing the PCR amplification product to obtain a base sequence, comparing the base sequence with the acrossocheilus fasciatus mitochondrial molecular marker, and identifying the water system source of the acrossocheilus fasciatus to be identified according to the specific site of the acrossocheilus fasciatus. According to the method, acrossocheilus fasciatus geographical populations distributed in different water systems are distinguished from gene features, and technical guarantee is provided for acrossocheilus fasciatus proliferation and releasing parent sources and offspring seed traceability.

The invention belongs to the technical field of DNA extraction, and particularly relates to a DNA rapid magnetic solid phase extraction method based on a magnetic functionalized nano material, which comprises the following steps: 1) preparing Fe3O4 NPs; (2) preparing Fe3O4 (at) PDA NPs; (3) preparing a magnetic UIO-66 (Zr) metal organic framework nano material; 4, DNA in the biological tissue is extracted.The prepared MUIO-66-COOH has a better magnetic solid phase adsorption extraction effect on DNA sodium salt, the magnetic solid phase DNA extraction method based on the MUIO-66-COOH has good precision, repeatability and stability, and the MUIO-66-COOH material has good recoverability and regeneration performance, so that the method can be used for efficiently and rapidly extracting DNA in a biological tissue sample.

1. Name of the product in this design: Reagent Kit. 2. Purpose of this design: Packaging box for packaging DNA extraction and testing reagents. 3. The key design elements of this product are: pattern, shape, and their combination. 4. The image or photograph that best illustrates the key design points: Design 1 3D view. 5. Design 1 is designated as the basic design.

The utility model relates to a DNA (deoxyribonucleic acid) extraction and DNA methylation conversion and purification device adopting a paramagnetic particle method. Comprising a machine body, an extraction box body which is fixedly arranged on the machine body and is internally provided with at least one extraction bin group, a conversion box body which is fixedly arranged on the machine body and is internally provided with at least one conversion bin group, a magnetic attraction assembly which adsorbs DNA through magnetic beads, and a driving assembly which is fixedly arranged on the machine body and is connected with the magnetic attraction assembly, the plurality of temperature assemblies are connected with the extraction box body and the conversion box body in a one-to-one correspondence manner, and the extraction bin group comprises a cracking bin chamber, a washing bin chamber, a magnetic bead bin chamber and an elution bin chamber which are sequentially arranged in the extraction box body; the transformation bin group comprises a transformation bin chamber, a desulfurization bin chamber, a washing bin chamber, a magnetic bead bin chamber and an elution bin chamber which are sequentially arranged in the transformation box body, the integrated operation of DNA extraction, DNA methylation transformation and purification can be automatically completed, purified DNA can be conveniently obtained, and instability and pollution in the manual operation process are reduced.

The invention relates to a water environment MST primer performance evaluation method, device and equipment and a medium. The method comprises the following steps: acquiring a water environment sample; the water environment sample comprises a target pollution sample and a non-target microorganism sample; performing DNA extraction on the water environment sample, and performing concentration standardization treatment on the water environment sample to obtain a positive sample library and a negative sample library; respectively carrying out amplification reaction on a target primer and the positive sample library and the negative sample library to obtain amplification curve data and melting curve data; based on the melting curve data and the amplification curve data, obtaining a sensitivity index and a specificity index corresponding to the target primer; obtaining a primer performance result based on the sensitivity index, the specificity index and a circulation threshold value in the amplification curve data; the primer performance result is used for representing whether the primer performance reaches the standard or not. By adopting the method, the accuracy of primer performance evaluation can be improved, and the reliability of water environment MST detection is guaranteed.

The application discloses a kind of fluorescence colorimetric double-mode detection streptococcus agalactiae without aerosol pollution method, comprising the following steps: (1) strain culture and DNA extraction;(2) the combination of UDG and LAMP;(3) UDG-LAMP-CRISPR fluorescence mode detection system;(4) colorimetric mode detection system: read image RGB value in G value, G value significantly increases, and it is positive result.This patent innovatively combines UDG, LAMP, CRISPR and G4, introduces connection probe to realize "fluorescence-colorimetric" single system double-mode detection.The two detection modes of the detection platform can be freely split or combined, and can meet the needs of different detection environments.By combining UDG system and LAMP technology, aerosol pollution possibly carried in amplification process is avoided.Connection probe is introduced, CRISPR fluorescence detection is combined with G4 colorimetric detection, a detection system with "fluorescence-colorimetric" two detection modes is established, which is used for quantitative identification and cross verification of streptococcus agalactiae.

Portable Capsule-Based DNA Extraction Device

The invention discloses an efficient extraction method of blue fox sperm DNA, and belongs to the technical field of molecular biology. Aiming at the structural characteristics that chromatin of the blue fox sperms is highly condensed, histone is replaced by protamine, and a stable nucleoprotein complex is formed through an intermolecular disulfide bond, the invention provides an efficient DNA extraction method suitable for the blue fox sperms. According to the method, based on the synergistic effect of three chemical reagents, namely SDS, PK and DTT, full lysis of the sperm cells of the blue foxes is achieved, and then efficient enrichment of sperm DNA of the blue foxes can be achieved by combining a conventional DNA extraction method. According to the efficient extraction method of the blue fox sperm DNA provided by the invention, the yield and integrity of the blue fox sperm DNA are remarkably improved, and the limitation that sperm chromatin is difficult to effectively lyse by a conventional method is overcome; the method provides reliable technical support for research on genetic diversity evaluation, population management, molecular breeding and the like of the blue foxes, and has the potential of popularization and application in other high-condensation sperm species.

The present invention relates to the technical field of identification of traditional Chinese medicine, specifically to an SSR fingerprint spectrum and identification method for garden ginseng, forest ginseng and wild ginseng. The SSR fingerprint spectrum of garden ginseng, forest ginseng and wild ginseng includes DNA extraction, PCR amplification, PCR product identification, and finally constructing the SSR fingerprint spectrum of garden ginseng, forest ginseng and wild ginseng; the SSR fingerprint identification method of garden ginseng, forest ginseng and wild ginseng includes the establishment of the SSR standard fingerprint spectrum of garden ginseng, forest ginseng and wild ginseng, the establishment of the sample DNA fingerprint spectrum, and the use of polymorphism comparison to identify the results. The fingerprint spectrum and identification method can be used for the identification of garden ginseng, forest ginseng and wild ginseng samples. The present invention can authenticate the authenticity of garden ginseng, forest ginseng and wild ginseng based on genomic characteristics, and the identification method has the advantages of being simple, rapid, reliable and highly reproducible.

The invention belongs to the technical field of biology, and particularly relates to an extracting solution for extracting DNA of sorghum seeds, a kit and an extracting method and application of the extracting solution. In order to solve the technical problems that the existing sorghum DNA extraction method is complicated, the extraction efficiency is low and long-term preservation cannot be realized, the invention provides a sorghum seed DNA extracting solution which comprises a DNA quick extracting solution and a cleaning solution, the rapid extracting solution contains Tris-HCl, NaCl, Tween-20 and EDTA (Ethylene Diamine Tetraacetic Acid); and the cleaning solution contains Tris-HCl and Tween-20 (polysorbate-20). By combining the extracting solution or the prepared kit with cellulose filter paper treated by NaCl, the DNA of the sorghum seeds can be rapidly extracted within 20 seconds, the extraction quality is high, the DNA of the sorghum seeds can be preserved for a short time, and meanwhile, the method has no inhibition effect on downstream PCR (Polymerase Chain Reaction). Therefore, the extracting solution, the kit and the extracting method can be quickly applied to fields or detection sites.

This invention relates to DNA extraction methods from soil and sediments with high humic substances content. The method focuses on the soil preparation phase; removing salts and cations with two different Tris-EDTA (TE) buffers, followed by precipitating humic substances with aluminum hydroxide solution and washing again with TE and cell lysis phase; urea is used as denaturing agents for the cell lysis phase and protein degradation phase while preventing the rediffusion of humic substances with CTAB. The purification can be performed following both the classic phenol chloroform method and using commercial DNA purification columns. This method is very accessible with operation time varying from 1h40 min to 2h30 min. The invention involves the use of TE and aluminum hydroxide solutions for soil preparation, and urea and CTAB as denaturing agents during cell lysis, in the specified concentrations, order, and incubation time, to prevent contamination of final DNA with humic substances

The invention discloses a method for counting the number of wild rock sheep based on a microsatellite molecular marker. The method comprises the following steps: preprocessing a rock sheep manure sample and extracting DNA (Deoxyribonucleic Acid); performing double PCR amplification on the microsatellite marker, wherein each group of microsatellite primers comprises two pairs of specific primers; carrying out electrophoresis separation on a PCR product by using 12% polyacrylamide gel, and reading the size of an allele fragment of each microsatellite site through GeneMapper software after silver staining and developing; analyzing the acquired allele data by using software, and counting the number of different wild rock sheep individuals in the region according to an individual identification standard; the individual identification standard is as follows: if the genotypes of all 10 microsatellite loci are completely the same, or only one allele of one locus is different, the same individual is determined. The method is particularly suitable for non-invasive individual identification, genetic relationship identification and population quantity statistics of plateau wild rock sheep populations, and provides technical support for rock sheep resource protection and dynamic monitoring.

The invention provides a full-process automatic all-in-one machine for DNA extraction, amplification and sequencing. The full-process automatic all-in-one machine comprises a cabin body, the cabin body is internally provided with a first space, and the first space is internally provided with a biological sample extraction workbench for realizing the whole process of DNA extraction, amplification and sequencing; the top of the cabin is provided with an air inlet duct, the side surface or / and the bottom of the cabin is provided with an air outlet duct, and the air inlet duct and the air outlet duct form negative pressure in the cabin; an amplification area is arranged on the biological sample extraction workbench, a cooling base used for reducing pollutants generated in the amplification process is arranged in the amplification area, and an air suction plate is arranged on the side face of the amplification area; the air suction plate is communicated with the air outlet channel, and pollutants generated in the amplification area are discharged out of the cabin body through the air outlet channel. Pollutants generated in the cabin body can be effectively prevented from facing three positions outside from gaps or cabin door lamps of the cabin body, so that pollution to the surrounding environment is avoided, and the safety of biological experiments is further improved. In addition, the integrated cabin is provided, and the requirement for the operation space of the biological experiment is effectively reduced.

According to the method for detecting and identifying the tea-seed oil by extracting DNA based on the membrane technology, the DNA of the tea-seed oil is separated, enriched and purified through a glass fiber membrane, the tea-seed oil is identified by combining a real-time fluorescent polymerase chain reaction (PCR) technology, and the method specifically comprises the steps of n-hexane fat dissolution, water phase separation and extraction, thermal cracking, purification, membrane method DNA extraction and real-time fluorescent PCR detection. According to the pretreatment method for extracting the DNA in the tea-seed oil based on the membrane technology, the real attribute of the tea-seed oil is rapidly identified through qualitative detection of endogenous genes by PCR (Polymerase Chain Reaction) detection; according to the method, trace DNA can be extracted from large-volume tea-seed oil, so that the requirements of a PCR detection method are met, and the problem that DNA in the tea-seed oil is seriously degraded, is damaged into small molecules and cannot be efficiently extracted can be well solved; nucleic acid molecules are effectively purified and separated through the glass fiber membrane, the detection cost is reduced, and the detection period is shortened.

The application discloses a plant genome DNA extraction reagent, which comprises Buffer 1 lysate, Buffer 2 precipitate and Buffer 3 combination liquid; the Buffer 1 lysate comprises sodium chloride, PVP40, sodium metabisulfite and SDS; wherein the concentration of the sodium chloride is 250-1200 mM; the mass percentage of the PVP40 is 0.5%-3%; the concentration of the sodium metabisulfite is 50-150 mM; and the mass percentage of the SDS is 0.7%-2.5%; the Buffer 2 precipitate comprises acetate; wherein the concentration of the acetate is 2.2-4.3 M; and the Buffer 3 combination liquid comprises guanidine hydrochloride; wherein the concentration of the guanidine hydrochloride is 0.5-2.5 M. The extraction reagent does not contain organic solvents such as phenol and chloroform, and can improve the removal rate of polyphenols.

The present invention belongs to the field of microbiology and genetics technology, and in particular relates to a primer set and typing detection method and kit for detecting drug-resistant enzyme genes of bacterial strains, wherein the primer set is an amplification primer for β-lactamase-resistant genes and carbapenemase-resistant genes, wherein the β-lactamase-resistant genes include SHV, TEM, and CTX-M, and the carbapenemase-resistant genes include NDM, OXA-23, and KPC. PCR amplification is performed according to specific enzyme typing primers, and genomic DNA is extracted using a magnetic bead method to improve the efficiency and purity of DNA extraction, thereby optimizing the accuracy and efficiency of PCR. The specific enzyme typing results are confirmed by comparison with the NCBI database through agarose gel electrophoresis and subsequent sequencing analysis. The experimental process is simplified, the cost is reduced, and a scientific basis is provided for the screening and clinical use of new β-lactamase and carbapenemase inhibitors, with the advantages of high efficiency, accuracy, and cost-effectiveness.

The invention discloses a method for detecting active watermelon bacterial fruit blotch bacteria by a PMA-PCR (Polymerase Chain Reaction) technology, which comprises the following steps: (1) collection and suspension of bacteria in a sample to be detected: suspending the sample with normal saline to obtain a bacterial suspension; (2) PMA treatment: adding propyl azide bromide (PMA) into the bacterial suspension; (3) blue light crosslinking: irradiating the bacterial suspension treated by the PMA under blue light to activate the covalent binding of the PMA and dead bacteria DNA (Deoxyribose Nucleic Acid); (4) DNA extraction: centrifugally collecting thalli, and extracting DNA by adopting a kit method; (5) PCR amplification: carrying out amplification by using a specific primer pair; and (6) result judgment: detecting an amplification product through agarose gel electrophoresis, wherein a target strip appears in an active bacterium sample, and no strip exists in a dead bacterium sample. The invention establishes a rapid, convenient, accurate and efficient method for detecting the active watermelon bacterial fruit blotch, so as to improve the disease detection capability and management efficiency.