29 results about "Microbial DNA" patented technology

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 belongs to the technical field of molecular biology, and particularly relates to a pathogenic microorganism nucleic acid co-extraction kit, an extraction method and application. The invention provides a lysis solution for nucleic acid co-extraction of pathogenic microorganisms. The lysis solution comprises a first guanidine salt, an ionic surfactant, a first sodium salt and a first nonionic surfactant. According to the splitting system, guanidine salt is adopted and matched with a certain content of sodium salt and a surfactant, a sample is split and precipitated under the grinding condition, nucleic acid cannot be damaged in the crushing process, and pathogenic microorganism DNA and RNA with high concentration and complete fragments can be extracted.

The invention discloses a method and system for synchronously detecting host chromatin openness and in-vivo microbiome based on transposase, and belongs to the technical field of biological sequencing data analysis. According to the method, transposase is used for selectively fragmenting an open chromatin region of a host, and a microbial genome is almost randomly cut, so that synchronous enrichment of host and microbial DNA is realized; after high-throughput sequencing library construction and double-end sequencing, sequencing data is split into host source and non-host source reads through bioinformatics analysis, host chromatin state and microorganism composition are analyzed respectively, and a microorganism-host epigenetic regulation network is constructed; the invention further provides a matched DNA sequencing library and an analysis system, multi-scene research of infectious diseases, intestinal microecology, tumor microenvironment and the like is supported, a public database can be reanalyzed, and potential microbial interaction signals are mined. According to the method, the host-microorganism interaction research efficiency is remarkably improved, and a high-sensitivity and integrated technical scheme is provided for epigenetic regulation mechanism analysis.

A portable extraction device consisting of a microfluidic cartridge, a magnetic bead-based cleaning system and an electronic control unit, configured for automated extraction of microbial DNA from soil samples.

The invention belongs to the technical field of petroleum geology engineering, and particularly provides an analysis method for microorganism DNA sequencing and application of the analysis method in stratigraphic division. The analysis method comprises the following steps that in the drilling process, rock debris is sampled from a vibrating screen at a wellhead, and rock debris samples are collected once every 0.5 m so as to collect rock debris samples of different stratum depths; the collected rock debris is subjected to treatment and DNA sequencing, and the variety and abundance of strains contained in the rock debris are obtained; the strains are preliminarily screened, and the strains from the underground oil reservoir are reserved; the reserved strains are clustered, and the total length of the stratum where the rock debris is collected is preliminarily divided into a plurality of stratums according to the similarity and correlation of the strains in different stratum depths; and each stratum is finely divided into a plurality of small stratums by taking the characteristic strains in each stratum as main characteristics.

The present invention is directed to methods for confirming the identity of an item or product using a segment of DNA present in a killed microorganism as a tag. In some embodiments, a food compatible microorganism containing a target nucleotide sequences not present in the item to be tagged is selected to serve as a tag and an identity unique identifier and / or other metadata for of the item is then saved for that tag in a database. The microorganism is killed to create a tag with the target nucleotide sequence and the tag is added to the item. The identity of the item can later be confirmed by extracting the DNA in the item and amplifying the target nucleotide sequence or sequences using PCR techniques, sequencing each amplified target nucleotide sequence; and then comparing them to the nucleotide sequence saved for the target nucleotide sequence.

The invention relates to the technical field of biomedicine and microorganisms, and provides an intestinal flora excrement sample preserving fluid which comprises a buffer system, a chelating agent, a reductive anaerobic protective agent, a cell lysis-bacteriostatic agent and a biomacromolecular stabilizer, wherein the buffer system is a combination of a Tris-HCl buffer solution and sodium chloride, the chelating agent is ethylenediamine tetraacetic acid disodium salt, the reductive anaerobic protective agent is L-cysteine hydrochloride, the cell lysis-bacteriostatic agent is sodium lauroyl sarcosinate, and the biomacromolecular stabilizer is a combination of trehalose and glycerin. The invention also provides a preparation method of the preserving fluid. According to the invention, the microbial DNA and community structure in the excrement can be stably preserved at room temperature for a long time without a cold chain; meanwhile, the preservation solution can effectively protect DNA of strict anaerobic bacteria from oxidative degradation, flora composition is prevented from shifting, and the reliability of subsequent molecular detection and analysis is finally ensured.

The application discloses a multi-cancer early screening method based on fragmentomics and microbiomics characteristics and application thereof, and belongs to the technical field of biomedical detection. The method comprises the following steps: extracting fragmentomics characteristics of human cfDNA and microbiomics characteristics of plasma microbial DNA based on sequencing data; inputting the two types of characteristics into a first-stage classification model after fusion, and judging whether a subject is cancer positive; and further predicting the cancer type through a second-stage classification model for the positive result. The application combines fragmentomics and microbiomics characteristics, and is used for multi-cancer early screening. Through single non-invasive blood sampling, early screening and organization tracing of eight high-incidence cancers such as lung cancer, liver cancer and colorectal cancer can be realized. The method has high sensitivity and high specificity under low sequencing depth, is low in cost, wide in coverage, and suitable for large-scale population screening.

The invention discloses an ultraviolet and hydrogen peroxide synergistic sterilization method. According to the method, an ultraviolet light source is arranged, a microbial DNA structure is directly destroyed by utilizing ultraviolet rays to realize primary inactivation, and meanwhile, a hydrogen peroxide solution is excited to be decomposed to generate high-activity oxidizing substances such as hydroxyl free radicals and the like. Through a dual action mechanism of physical irradiation and chemical oxidation, a multi-dimensional synergistic sterilization system is formed, and various microorganisms with cellular structures, such as fungi, bacteria and spores, can be efficiently killed. Compared with a traditional single sterilization technology, the sterilization method has the application advantages that the sterilization blind area is eliminated through a multi-effect synergistic mechanism, and sterilization thoroughness is improved; a low-concentration hydrogen peroxide solution is adopted, so that the chemical residue risk is reduced. The method is realized through a special ultraviolet and hydrogen peroxide synergistic sterilization device, and is suitable for object surface treatment.

The utility model relates to the technical field of boards, and discloses a plasma antibacterial board which comprises a wood board base material, surface layer boards are arranged at the upper end and the lower end of the wood board base material, the surface layer boards are antibacterial boards processed through the plasma technology, the surface layer boards are hot-pressed on the wood board through high temperature and high pressure generated by ionized gas, and antibacterial factors are contained in the surface layer boards. After the antibacterial factors are catalyzed by high temperature and high pressure generated by ionized gas, quadruple antibacterial effective components including zinc ions, hydroxyl radicals, superoxide ions and hydrogen peroxide are formed. The antibacterial effective components can destroy the protein structure of bacteria and interfere the DNA synthesis of microorganisms, so that the efficient antibacterial effect is realized. The efficient and lasting antibacterial property is realized, and the bacterium breeding can be effectively inhibited.

The present invention is directed to methods for confirming the identity of an item or product using a segment of DNA present in a killed microorganism as a tag. In some embodiments, a food compatible microorganism containing a target nucleotide sequences not present in the item to be tagged is selected to serve as a tag and an identity unique identifier and / or other metadata for of the item is then saved for that tag in a database. The microorganism is killed to create a tag with the target nucleotide sequence and the tag is added to the item. The identity of the item can later be confirmed by extracting the DNA in the item and amplifying the target nucleotide sequence or sequences using PCR techniques, sequencing each amplified target nucleotide sequence; and then comparing them to the nucleotide sequence saved for the target nucleotide sequence.

A grade of a specific microbial disease in a biological sample of a subject is determined. In one example, masked microbial reference genomes are used to determine the amount of free DNA molecules corresponding to a particular microbial species associated with a particular microbial disease. The masking may remove an area shared with another. In another example technique, end motifs are used for free DNA fragments from a subject and from a particular microbial species. A correlation between a subject and an amount of a set of terminal sequence motifs of a particular microbial species may be determined. The two groups of quantities of positive subjects are substantially more correlated than negative subjects.

The invention discloses a gastric cancer risk assessment and diagnosis method based on gastric mucosa micro-ecological change, relates to the technical field of medical diagnosis and biology, and provides the gastric cancer risk assessment and diagnosis method based on the gastric mucosa micro-ecological change. The method comprises the following steps: collecting gastric mucosa tissues or gastric juice samples, extracting total DNA (deoxyribonucleic acid) of microorganisms, performing high-throughput sequencing by using an Illumina MiSeq platform, analyzing the diversity of the microorganisms, and identifying key flora. And inputting a random forest model by combining the concentration of metabolites such as short-chain fatty acid and clinical information, outputting a gastric cancer risk score, and dividing risk grades. The model is trained based on a large number of samples, and high accuracy is ensured. High-risk persons suggest gastroscope precise examination within three months, and personalized intervention suggestions are provided, so that early warning and precise health management are realized.

Methods are provided for detecting disease in liquid biopsy samples by identifying microbial rRNA. The methods include isolating nucleic acids from extracellular vesicles, treating the nucleic acids with DNase to remove DNA including human cfDNA, mcfDNA, and contaminating microbial DNA, and then amplifying a region of a microbial phylogenetic rRNA. To further reduce contaminating DNA, the amplifying step can include reverse transcription to translate the phylogenetic region into cDNA and extend the cDNA ends with a T7 promoter and sequencing adaptors. The translated region is amplified with primers to the extended ends of the cDNA and sequenced. The resulting phylogenetic microbial sequences are useful for diagnosis and monitoring of diseases including cancer. The method can further include amplification of other human or bacterial DNA or RNA sequences to augment disease diagnosis. The method results in a very low contribution of contaminating microbial DNA and RNA to the overall microbial signature.

The present disclosure provides systems and methods for detection of microbial DNA and RNA. In an aspect, a system or method for detection of microbial DNA or RNA comprises metagenomic next-generation sequencing of clinical samples (e.g., cerebral spinal fluid) to detect the microbial composition of the sample to diagnose a microbial infection (e.g., of the central nervous system).

The present application relates to the technical field of ship biological killing, in particular to a UV and ultrasonic synergistic killing control method and device for ballast water treatment, which is constructed with a central control unit as the core, integrates a synergistic control device of multiple sensors such as flow, turbidity, microbial content, ultraviolet intensity and temperature, and realizes real-time monitoring of the ballast water quality and the equipment operation state. The ultrasonic generating unit and the ultraviolet disinfection unit work synergistically under the precise regulation and control of the central control unit, utilize the ultrasonic cavitation, mechanical and thermal effects, combine the ultraviolet damage to the microbial DNA, efficiently kill various microorganisms and remove the scale on the ultraviolet lamp. The present application improves the microbial killing efficiency, prolongs the service life of the ultraviolet lamp, reduces the equipment maintenance cost and energy consumption, and ensures that the ballast water discharge meets the environmental protection standards.

The invention discloses disinfection equipment for a root canal file. The disinfection equipment consists of a box body, a cleaning device, a disinfection and drying device and an intelligent control module, the box body is divided into an upper layer and a lower layer, and the cleaning device cleans the root canal file at 360 degrees through a nozzle. And the lower layer is a disinfecting and drying device which comprises a chemical disinfecting device, a high-temperature steam disinfecting device, an ultraviolet disinfecting device and a drying device. The chemical disinfection device adopts an environment-friendly disinfectant; the high-temperature steam sterilization device utilizes an efficient electric heating pipe and a steam-water separation device to generate steam to kill microorganisms; the ultraviolet form device destroys microorganism DNA by using a high-efficiency ultraviolet lamp tube; the drying device forms hot air circulation to evaporate moisture through a fan and a hot-air blower. The intelligent control module can automatically select programs and parameters, monitor the operation state, give an alarm and record data. The equipment integrates various technologies and realizes comprehensive, efficient and safe disinfection of the root canal file.

This invention belongs to the field of plant rhizosphere research and discloses a method for determining the regulatory effect of exogenous selenium application on the rhizosphere microecology of Amorphophallus bulbifera. The method includes: experimental design and sample collection; soil microbial DNA extraction and amplicon sequencing; and quality control and analysis of the microbiome data. This invention utilizes Illumina MiSeq high-throughput sequencing technology to analyze the changes in the composition and diversity of rhizosphere soil bacteria communities in Amorphophallus bulbifera under exogenous selenium treatment, and further analyzes the functions of related communities through PICRUSt functional prediction analysis. The conclusions are as follows: exogenous selenium treatment induces significant changes in the rhizosphere soil bacterial community of Amorphophallus bulbifera, which is beneficial for creating a stable and more diverse rhizosphere microbial community structure.

The invention belongs to the technical field of biology, and particularly relates to a method for extracting metagenomes of microorganisms in a mud-water mixture. Comprising the following steps: S1, mud-water sample separation: separating mud and a water body part in mud-water; s2, extraction of metagenomes in the soil: extracting the metagenomes in the soil; s3, water body sample separation: separating microorganisms from free DNA in the water body; s4, extracting metagenomes of thallus samples in water, and extracting metagenomes of microorganisms in the water; and S5, extracting metagenome of the free DNA sample in water, and extracting free DNA in the water. According to the method disclosed by the invention, almost all microorganism DNAs can be effectively recovered from the mud-water mixture, and the DNA loss caused by sample separation is reduced. The DNA recovery rate is improved through optimized splitting decomposition and purification steps, and it is ensured that a high-quality DNA sample can be obtained even under extreme conditions.

The application discloses a method and system for predicting gene expression regulated by microbial DNA methylation, comprising genome sequencing, obtaining genome assembly data and DNA methylation modification signals; genome assembly data annotation, obtaining transcription regulation regions and gene coding regions; selecting methylation sites from the DNA methylation modification signals, obtaining a first position overlapping region of the methylation sites and the transcription regulation regions and a second position overlapping region of the methylation sites and the gene coding regions; and predicting the relationship between the methylation sites and the genes according to the two position overlapping regions. The application establishes a one-to-one correspondence between genes and DNA methylation by using SMRT sequencing technology, can obtain genes in the whole genome that may be regulated by DNA methylation, thereby reducing the experimental time, narrowing the range of genes regulated by DNA methylation, and improving the interpretation of the regulation mode of microbial DNA methylation in a high-throughput manner.

The present invention relates to systems and methods for well planning and management in a field to maximize the production of hydrocarbons. Specifically, the present invention involves identifying microbial compositions (including phenotypic microbial compositions) of solid samples, such as cuttings or core samples, or fluid samples, such as produced or injected fluids and determining well communication or fracture height (frac height). With well communication and / or frac height analysis using microbial compositions, the operator can better plan the placement and geometry of wells in a field to maximize production and reduce the number of wells that need to be drilled to maximize that production.

The present invention provides improved methods and kits for isolating microbial nucleic acids, preferably microbial DNA, from a tissue sample comprising a eukaryotic tissue, said method comprising the steps of: a) treating the tissue sample with a collagenase and a neutral protease that is different from the collagenase, b) treating the sample of step a) with at least one nuclease, and c) processing the sample of step b) to isolate microbial nucleic acids, preferably microbial DNA. The method significantly improves depletion of host cell DNA.

The invention discloses a host DNA removal method based on magnetic separation. According to the method, human cells in a to-be-detected sample are specifically adsorbed through streptavidin magnetic beads, complete cells in the sample are pulled out, free DNA in the sample is removed through nuclease, a purified microorganism DNA solution is obtained, and finally genome DNA is extracted and can be used for related sequencing and research. According to the DNA solution obtained by the method disclosed by the invention, the content of host DNA is greatly reduced, so that the sequencing accuracy is improved.

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 invention relates to a preparation method of a natural mineral-based ternary synergistic antibacterial composite material. The antibacterial composite material takes natural clinoptilolite as a carrier to synergistically load a rare earth catalytic component and bimetallic antibacterial ions; the preparation method comprises the following specific preparation steps in sequence: pickling-thermal activation pretreatment of natural clinoptilolite, loading of rare earth catalytic components, synergistic loading of bimetal ions, optional surface stabilization treatment and post-treatment. Through the ternary synergistic effect of rare earth catalytic components and bimetallic ions, an adsorption-slow release-catalysis three-in-one antibacterial system is constructed, a zeolite carrier adsorbs microorganisms, the rare earth components catalyze and destroy microbial cell membranes, Ag < + > and Zn < 2 + > respectively destroy microbial DNA and enzyme activity, and through the synergistic effect of the zeolite carrier, the rare earth components and the Ag < + > and the Zn < 2 + >, the material has a good antibacterial effect on Gram-positive bacteria, Gram-negative bacteria, Gram-positive bacteria, Gram-negative bacteria, Gram-negative bacteria and Gram-negative bacteria. The antibacterial effect is lasting, the release is controllable, and the burst release phenomenon is avoided; meanwhile, the synergistic effect of the rare earth oxide and the bimetal ions can prevent the microorganisms from generating drug resistance.

This invention discloses an anti-biofouling UV irradiation system based on an online marine environmental observation system, belonging to the field of marine environmental monitoring equipment technology. Addressing the problem of biofouling on marine observation equipment, this invention employs modular, rotatable, stackable UV LED modules, a waterproof and pressure-resistant housing, a high-transparency quartz outer cover, standardized pluggable interfaces, and a low-power intelligent control unit. It uses 254nm UV light to disrupt microbial DNA replication and block biofouling. The LED modules can be adjusted 360° and freely assembled, adapting to various irregularly shaped sensors. Combined with an MSP430 processor, it achieves intermittent frequency conversion and time-series linkage light control. This invention provides purely physical antifouling without secondary pollution, has wide adaptability, low power consumption, long-lasting effectiveness, and stable operation. It can extend the equipment maintenance cycle and ensure the accuracy of observation data, making it suitable for in-situ antifouling of underwater observation facilities such as buoys, submersible moorings, and seabed observation networks.

The invention discloses an identification method for a water body odor substance 2-MIB generation mechanism, and the method comprises the steps: selecting a target water body, and obtaining water body parameters; collecting multi-source data according to a first sampling period; the method comprises the following steps: extracting total DNA (deoxyribonucleic acid) of microorganisms in a target water body, identifying algae community composition through a high-throughput sequencing technology, and screening blue-green algae types generating odor substances; detecting the copy number and / or expression quantity of the blue-green algae by taking the mic olfactory gene as a target and adopting a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) technology, and constructing olfactory change trends at different time points; performing correlation analysis on the algae composition, the mic gene abundance, the environmental factors and the concentration of the odor substances, and constructing an odor production mechanism recognition model; and identifying the high-risk olfactory population and the key driving factors thereof based on the olfactory mechanism identification model, and outputting an olfactory mechanism identification result. According to the method, on the basis of a set of sampling data, high-throughput sequencing and molecular quantitative detection are combined, so that smelling populations and key driving factors thereof are quickly locked, and occurrence of water body peculiar smell events is controlled from the source.

The application discloses the technical field of petroleum geology engineering and specifically provides a microbial DNA sequencing analysis method and application thereof in formation division, which comprises the following steps: during drilling, sampling cuttings from a shaker at a wellhead, and collecting a cuttings sample every 0.5 meters to collect cuttings samples at different formation depths; treating the collected cuttings and performing DNA sequencing to obtain the types and abundance of bacteria contained in the cuttings; preliminarily screening the bacteria, and retaining bacteria from an underground oil reservoir; clustering the retained bacteria, and preliminarily dividing the total length of the formation from which the cuttings are collected into multiple formations according to the similarity and correlation of the bacteria at different formation depths; and finely dividing each formation into multiple small layers by taking characteristic bacteria in each formation as main characteristics.

The invention provides a method for detecting the influence of microplastics and triclosan on the structure and function of a sediment microbial community. The method comprises the following specific steps: firstly, collecting a sediment sample in a specified sampling area; then adding the micro-plastic particles and a triclosan solution into the sediment sample, and incubating; dNA of the microorganisms is extracted, and 16S rRNA sequencing is carried out; and finally, analyzing the diversity of the microbial community and evaluating the structure and function of the microbial community according to the sequencing data. The comprehensive influence of micro-plastic and triclosan combined pollution on the structure and function of the sediment microbial community is systematically analyzed for the first time, the efficiency and accuracy of sediment microbial community detection are effectively improved by adopting a 16S rRNA detection technology, and a reliable basis is provided for formulating an environment protection strategy, optimizing pollution control and promoting ecological restoration engineering.