31 results about "DNA extraction" patented technology

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.

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 application provides a DNA extraction method and application thereof. The DNA extraction method comprises the following steps: crushing cells of a sample to obtain a first mixture; mixing the first mixture with a hexammine cobalt (III) chloride solution and an adsorption carrier, performing first incubation, and obtaining a first incubation product; washing and dissolving the first incubation product to obtain DNA of the sample. The method can solve the problems of the prior art, such as complexity of the DNA extraction method and low quality of the obtained DNA, and is suitable for the field of DNA extraction.

The application discloses a kind of based on toxigenic and odor-producing gene toxigenic and odor-producing cyanobacteria field rapid monitoring system and method, a kind of based on toxigenic and odor-producing gene toxigenic and odor-producing cyanobacteria field rapid monitoring system, including water sample algal automatic enrichment module, algal cell rapid lysis and DNA extraction tube, isothermal / normal temperature PCR module, palm fluorescence detector, qualitative and quantitative algorithm and toxigenic and odor-producing cyanobacteria risk warning grade calculation software, mobile phone APP software, each module is sequentially linked.The application realizes the integrated detection of toxigenic and odor-producing cyanobacteria quickly, accurately and conveniently, solves the problem that existing technology cannot accurately distinguish between toxigenic and non-toxigenic cyanobacteria and is difficult to scientifically assess the health risk of water body, while overcoming the limitations of traditional monitoring technology, such as laboratory detection, large equipment, complex operation, long detection period, etc., and meeting the timeliness, convenience and ease of operation requirements of field scenes such as grassroots monitoring, outdoor inspection and emergency disposal of sudden water bloom.

This invention relates to the field of ecological conservation technology, specifically to a method and system for screening species that preserve ecological memory from discarded polypropylene masks. The method includes: obtaining baseline and transfer samples of biofilm from discarded polypropylene masks; obtaining a relative abundance matrix after DNA extraction, high-throughput sequencing, and quality control standardization; and extracting the top-ranking microorganisms from the transfer group to construct an independent variable feature matrix. Ecological memory retention is calculated based on the Bray-Curtis distance between the transfer group and the homologous baseline group. A random forest model is trained using the feature matrix, and key microorganisms are screened based on the error changes after shuffling the abundance. Ecological memory-preserving species are then screened based on the Spearman correlation between their abundance and retention. Finally, the intersection of ecological memory-preserving species, network key species, and neutral community key species is defined as the super-core species. This invention improves the accuracy of screening ecological memory-preserving species.

The application provides a DNA extraction method for leaves of a plant of the genus Catalpa, and relates to the technical field of DNA extraction of plant leaves. The method is based on the CTAB method, and a lysis solution is optimized so that the extraction method is suitable for the plant of the genus Catalpa. The DNA extraction method for leaves of the plant of the genus Catalpa has the beneficial effect of high extraction rate.

1. The name of the design product: DNA integrated nucleic acid extraction workstation. 2. The use of the design product: for DNA extraction. 3. The design points of the design product: in shape. 4. The picture or photo that best indicates the design points: perspective view.

This invention belongs to the field of biotechnology, specifically relating to a non-harmful DNA extraction method for *Scylla serrata* (scuttica crab) and its application. This invention successfully achieves non-harmful DNA extraction from *Scylla serrata* by collecting its swimming leg setae, significantly reducing the impact on the crab's activity and ensuring its survival after sampling. The extraction method of this invention is non-lethal to *Scylla serrata* and possesses good repeatability and high accuracy. Extracting DNA from *Scylla serrata* using its swimming leg setae ensures that the crab's activity remains unchanged, allowing it to survive and perform its life activities normally. Obtaining the DNA sequence of *Scylla serrata* and performing gene markers provides a foundation for genomic selection breeding, facilitating parent selection and offspring breeding in mud crab genetics.

This invention discloses a water sampling bag for centrifugally separating environmental DNA from sediment and its usage method. The bag includes a threaded sealing cap at the top, a hanging handle on the outside, a water sample collection interface with a threaded cap embedded on one side of the upper end, and a bottom bladder and a drain pipe with a water-stop clamp at the lower end. The side of the bag has graduations, and the inner side of the upper end has a connection point and a bag opening. The threaded sealing cap is threaded into the bag opening. The hanging handle is adapted to the bag opening via a limiting shaft and a combination ring. The water-stop clamp includes a clamping plate and an adjusting screw. The method utilizes this water sampling bag, including two water sample injection methods. After centrifugation at 500-2000 rpm, sediment is concentrated in the bottom bladder. The supernatant is poured out and filtered through a membrane to collect environmental DNA from the water. The sediment in the bottom bladder is dissolved in anhydrous ethanol and collected for preservation or DNA extraction. This invention has a reasonable structure, reliable sealing, avoids contamination, is adaptable to different water sample collection methods, and efficiently separates water samples from environmental DNA samples.

The application discloses a Super-long fragment DNA extraction kit and application. The Super-long fragment DNA extraction kit is universal, can solve the DNA super-long fragment extraction of culture cells, cryopreserved cells, fresh animal tissues, cryopreserved animal tissues, common crops, polysaccharide and polyphenol plants and part of silica gel dry plant samples and the like, can guarantee the purity under the condition that the length reaches 500kb-1Mb, and is suitable for long read length sequencing with high requirements for the DNA fragment length and the DNA purity.

The application discloses a kind of efficient collection and extraction method and application of air eDNA.The method uses glass fiber filter as sampling filter, uses medium-flow particulate matter sampler and TSP sampling head to carry out sampling, uses dry ice to carry out sample preservation transportation, uses PBS buffer and low-temperature low-speed centrifugal elution DNA, uses PES filter to enrich and precipitate particulate matter, and PES filter is cut into small pieces to improve DNA extraction efficiency.The environmental DNA extracted by the application has higher concentration and good integrity, and the DNA extracted by the method can obtain good amplification results using universal primers of different species for amplification, which is convenient for subsequent eDNA macro-barcode technology to analyze species diversity.

The application discloses a kind of SSR molecular marker primer for identifying Cymbidium varieties, and its application, the Cymbidium includes Cymbidium kanran, Cymbidium ensifolium, Cymbidium goeringii, Cymbidium sinense, Cymbidium devonianum, Cymbidium lancilobatum, Cymbidium kanran, hybrid orchid and hybrid Cymbidium. The SSR molecular marker includes SSR1-SSR23, and the primer sequence is shown as SEQ ID NO.1-SEQ ID NO.46 in the sequence table. The screening method of the SSR molecular marker primer is also included, which comprises the following steps of total DNA extraction of Cymbidium, SSR site selection and primer design based on Cymbidium reference genome, PCR amplification, polyacrylamide gel electrophoresis primary screening and ABI3730XL gene analyzer secondary screening. The application also provides a composition comprising the primer and its application in Cymbidium genetic relationship analysis, DUS specificity test and molecular marker assisted breeding. The SSR molecular marker primer provided by the application has the characteristics of high polymorphism, large quantity, low cost, easy reading, and is not limited by environment, and can quickly and accurately construct the DNA fingerprint of Cymbidium varieties.

The utility model discloses a rhinopithecus roxellana excrement sample storage device for DNA extraction relates to rhinopithecus roxellana technical field, including box, be provided with circular cavity in the box, be provided with circular column in rotation in the circular cavity, three placement cavities of circular array distribution are seted up to the circular column outside, the box mouth of penetrating type is shaped to one side of circular cavity, the purification mechanism that purifies the air in the placement cavity inside is provided with in the box, the circular column outer wall and circular cavity inner wall contact, this rhinopithecus roxellana excrement sample storage device for DNA extraction, through setting rotatable circular column, so make two placement cavities in circular column can always keep sealed state, so reduce the excessive peculiar smell of box interior to discharge, also can increase the sealing property of the storage jar of the storage in the box, and thus effectively improve the practicality of rhinopithecus roxellana excrement sample storage device for DNA extraction, and reduce the peculiar smell emission too much and influence the working environment.

The application discloses a triplex forming oligonucleotide fluorescent probe and a detection method for detecting mycoplasma by targeting P1 adhesion protein genes, and belongs to the technical field of microbial molecular detection. The fluorescent probe is a triplex forming oligonucleotide (TFO), targets a specific conservative sequence of mycoplasma P1 adhesion protein genes, the 5' end of the fluorescent probe is modified with a fluorescent reporter group, the 3' end of the fluorescent probe is modified with a fluorescent quenching group, the fluorescent probe can form a stable triplex DNA complex with the target sequence through Hoogsteen base pairing and generate a fluorescent signal. The detection method comprises four core steps of sample pretreatment and DNA extraction, fluorescent probe incubation, fluorescent signal detection and result judgment, and does not need PCR amplification, and is simple in operation and rapid in detection. The probe has high specificity and high sensitivity, the detection lower limit can reach 1 copy / muL, is suitable for mycoplasma detection of various types of samples, and can be widely applied to scenes such as clinical diagnosis, biological pharmaceutical quality control and livestock breeding quarantine.

The subject invention describes a new method for the detection of root-knot nematodes (Meloidogyne spp.) at the egg stage isolated from soil. The described procedure enables the isolation of the eggs of root-knot nematode species directly from the soil and their subsequent use for their detection and identification without DNA extraction. In the procedure, an enzyme cocktail is used to release the DNA content from the material obtained by a specific purification procedure of the soil being tested and put into a real time PCR reaction, which detects the presence of nematodes of the Meloidogyne species. The procedure described here can be easily modified for the identification of other types of parasitic nematodes such as Globodera spp, Heterodera spp ili Pratylenchus spp.

The application provides an environmental DNA extraction method and device. The environmental DNA extraction device comprises an upper seat body, a lower seat body and a detachable membrane assembly. The upper seat body is a column with a sealed upper end and an open lower end. Tangential flow water inlets and tangential flow water outlets are arranged on the two side walls of the upper seat body. The upper end of the lower seat body is open, and the lower end of the lower seat body is provided with a filtrate outlet. The lower end of the upper seat body is connected to the upper end of the detachable membrane assembly, and the upper end of the lower seat body is connected to the lower end of the detachable membrane assembly. The detachable membrane assembly is arranged between the upper seat body and the lower seat body. Environmental water flows into the detachable membrane assembly through the tangential flow water inlets. The detachable membrane assembly filters and adsorbs the environmental water to adsorb environmental DNA in the environmental water on the detachable membrane assembly, and the remaining filtrate flows out through the filtrate outlet of the lower seat body. The extraction process can be simplified, the time consumption can be reduced, the risk of DNA degradation and sample pollution can be avoided, and the DNA extraction quality can be improved.

The application discloses a kind of InDel marker fingerprint of wild Phyllostachys annamica strain, belong to red-to bamboo cane strain detection technical field, solve the problem that existing technical means cannot accurately identify wild red-to bamboo cane strain, the construction of the InDel marker fingerprint of the wild red-to bamboo cane is by genome DNA extraction, construct amplification system and amplify genome DNA, and the electrophoresis detection of amplification product, to obtain the InDel marker fingerprint of red-to bamboo cane strain, this fingerprint is used as standard atlas, the electrophoresis band in atlas is marked as 1; Lower band is marked as-1; Two bands are marked as 2;The InDel marker fingerprint of the wild red-to bamboo cane strain obtained by the above numbering step is-1, 2, 2, 2, -1, 1, -1, 1, 2, 2, subsequent unknown red-to bamboo cane strain is compared by constructing fingerprint, and the obtained fingerprint is compared with standard atlas.

The application discloses a crab meat DNA extraction kit and an extraction method, and belongs to the field of molecular biology. The kit comprises a lysis solution, and the lysis solution comprises SDS, protease K and RNase A. The kit is simple in components, easy to prepare, low in cost and free of toxicity. The extraction method is simple and easy to operate, and crab meat DNA can be quickly extracted, thereby promoting the high-quality and sustainable development of the aquaculture industry.

The application belongs to the technical field of DNA extraction, and particularly relates to a field rapid extraction method of DNA in plant bacterial disease lesion, which comprises the following steps: mixing plant disease leaf lesion tissue and a lysis solution, and heating in a water bath in a thermos cup to complete nucleic acid release; the lysis solution comprises 0.05-0.2 mol / L NaOH; preferably, the lysis solution comprises 0.05-0.2 mol / L NaOH, 0.01% SDS and 0.1 mmol / L EDTA. The DNA extraction method established by the application does not need precise instruments and toxic reagents, is simple to operate, short in time consumption, and can realize field rapid extraction of plant lesion bacterial DNA, thereby providing technical support for early diagnosis and accurate prevention and control of plant bacterial diseases in the field.

The present application belongs to the technical field of molecular markers, and particularly relates to a tartary buckwheat stalk strength molecular marker and application thereof in identifying tartary buckwheat stalk strength traits. The molecular marker provided by the present application is located at 35,829,660 bp on the first chromosome of tartary buckwheat, and the single nucleotide polymorphism is C / G; when the base is C, the tartary buckwheat exhibits the trait of strong stalk strength. Based on the molecular marker, the stalk strength trait of tartary buckwheat can be selected, and the identification of samples can be completed only by simple DNA extraction, PCR specific amplification and KASP genotyping detection, so that tartary buckwheat with strong stalk strength and tartary buckwheat with poor stalk strength can be distinguished, the tartary buckwheat stalk strength can be quickly screened and identified, the breeding efficiency of tartary buckwheat germplasm with strong stalk strength is improved, a basis is provided for the utilization of excellent allelic variations related to the tartary buckwheat stalk strength trait, and the breeding process is accelerated.

This invention discloses a contamination-proof centrifuge tube device for micro-DNA extraction, relating to the field of DNA extraction technology. The device includes a centrifuge tube body with a first threaded groove on its inner wall. A connecting mechanism is installed inside the centrifuge tube body through the first threaded groove. A third thread is fixedly connected to the outer wall of the centrifuge tube body. A second sealing mechanism is provided at the top of the connecting mechanism. Through the second sealing mechanism, the liquid silicone valve deforms during the insertion or removal of the dropper, effectively preventing contamination. The first sealing mechanism effectively blocks the second sealing mechanism, preventing liquid leakage during centrifugation and preventing bacteria from entering the centrifuge tube body and contaminating the liquid due to deformation of the liquid silicone valve during centrifugation.

This invention discloses a nanopore sequencing method for detecting full-length 16S rRNA amplicon in human semen samples. The method includes sample pretreatment and DNA extraction; nested PCR amplification using outer primers 16S-27F / 1492R and inner primers 16S-340F / 1390R to obtain target long amplicon lengths of 1.0kb to 1.1kb; construction of a nanopore library and single-molecule long-read sequencing; deep filtering of the raw data with a quality score Q≥20 and read lengths between 1000bp and 1700bp; and finally, feature clustering and taxonomic annotation. This invention addresses the characteristics of low biomass and high host DNA background in human semen samples by employing innovative specific nested primer design and customized data cleaning strategies. This effectively avoids non-specific interference and significantly improves the classification resolution and data fidelity of the semen microbiome at the genus and species levels, providing a precise detection method for analyzing the reproductive microecology of infertile men.

FIELD: biotechnology.SUBSTANCE: used in diagnostic and research laboratories. A method for producing rod-shaped magnetite particles for isolating deoxyribonucleic acid (DNA) from whole blood is proposed, which includes producing Fe3O4 magnetite rods. At the first stage, particles of akagenite β-FeOOH are formed by hydrolysis of 0.57 M aqueous solution of FeCl3 at a temperature of 120–125 °C for 3 h. After completion of the reaction, the suspension is cooled to room temperature, the precipitate is separated by centrifugation at 10000 g for 10 min, washed with deionized water and ethanol and dried at 60 °C throughout the day. The resulting akagenite particles are dispersed in 20 ml of ethylene glycol using ultrasonic treatment at 100 W for 15 minutes. The resulting suspension is placed in an autoclave and subjected to hydrothermal treatment at 200 °C for 8 hours with the formation of the magnetite phase Fe3O4. Magnetite particles are separated with a magnet, washed successively with water and ethanol and dried at 60 °C within 24 hours. Then, a SiO2 shell is formed by dispersing a 50 mg sample of dried magnetite rods in 5 ml of deionized water. Dispersion is carried out in five cycles, each of which includes stirring at 300 rpm for 10 min and ultrasonic treatment at 45 W for 3 min. Isopropanol is added to the resulting suspension at a rate of 15 ml of isopropanol for every 5 ml of water, after which the mixture is stirred for 20 min at 300 rpm. Then, while stirring, 0.5 ml of a 25% NH4OH solution and 0.5 ml of tetraethoxysilane are added to the aqueous alcohol dispersion of particles for every 50 mg of rods. The formation of the SiO2 shell is carried out at room temperature for 20 hours with constant stirring. After incubation, the particles are purified by magnetic decantation with deionized water at least five times. The process of forming a silicon dioxide shell is repeated twice, and the purified rod-shaped particles of Fe3O4@SiO2 are dispersed in a 5% solution of polyethylene glycol with a molecular weight of 4000 Da. Fe3O4@SiO2-NH2 particles can also be obtained.EFFECT: increasinng the quantity and improving the purity of DNA isolated from whole blood by using magnetic particles with increased specific surface area and sorption capacity.2 cl, 12 dwg, 2 tbl, 3 ex

This invention discloses a method for systematically isolating and identifying Verticillium wilt of cotton, with the following steps: (1) disease sample collection and initial treatment; (2) strain culture: transfer the diseased tissue to a culture dish containing ampicillin PDA medium and culture it in a constant temperature incubator at 28°C for 3-5 days; (3) streak isolation and colony screening: place the culture dish under a microscope to directly observe the colony morphology. If the colony has the characteristic morphological features of Verticillium wilt of cotton, then the culture dish is confirmed to contain Verticillium wilt of cotton colonies; (4) single colony purification: from the culture dish containing Verticillium wilt of cotton colonies confirmed in step (3), pick a single colony with a sterile inoculation loop, transfer it to a new PDA medium for streak culture, and perform 2-3 consecutive passages for purification to obtain a purified strain; (5) DNA extraction and molecular identification; (6) pathogenicity verification inoculation; (7) re-isolation verification.

This invention provides a method and kit for extracting microbial DNA from marine sediments. The extraction method includes: a) washing the marine sediments with a washing buffer to obtain a washed sample; wherein the washing buffer includes spermidine and sodium pyrophosphate; b) lysing the microbial cells in the washed sample to obtain a lysed sample; c) recovering, screening, and purifying the DNA from the lysed sample to obtain microbial DNA. This method solves the problem of poor DNA extraction efficiency in existing technologies and is applicable to the field of DNA extraction.

The application discloses a triplex forming oligonucleotide fluorescent probe and a detection method for detecting mycoplasma by targeting CARDS toxin genes, and belongs to the technical field of microbial molecular detection. The fluorescent probe is a triplex forming oligonucleotide (TFO), which is targeted to a specific and conservative core sequence of mycoplasma CARDS toxin genes, and a fluorescent reporter group is modified at the 5' end, and a fluorescent quenching group is modified at the 3' end. The TFO can form a stable triplex DNA complex with the target sequence through Hoogsteen base pairing, and trigger the release of a fluorescent signal. The detection method comprises sample pretreatment and DNA extraction, fluorescent probe incubation, fluorescent signal detection and result judgment in sequence, and does not need a PCR amplification step, and the whole detection process can be completed within 45 minutes. The probe takes a functional conservative region of the CARDS toxin gene as a target, effectively avoids detection deviation caused by variation of surface antigen genes, has a specificity of 100%, and has a detection lower limit of 1 copy / muL, and is suitable for mycoplasma detection in multiple scenes such as clinical treatment, biological pharmaceuticals and livestock breeding.

The method for determining the sorting condition of plasma (105) has the following steps: measuring a blood sample by a blood state confirmation unit; calculating a converted elapsed days from a blood sampling date of the blood sample based on the measurement result; calculating a gDNA amount in the blood sample based on the converted elapsed days; and determining a plasma sorting condition based on the calculated gDNA amount. Thus, the method for determining the sorting condition of plasma, the plasma sorting device, and the DNA extraction device provided with the plasma sorting device, which can realize the plasma sorting process in a more appropriate condition compared with the past.