75 results about "Microbe DNA" patented technology

A method for identifying microbial DNA/RNA comprising providing a sample containing microbial DNA/RNA; enriching DNA/RNA in the sample by a PCR method wherein the DNA/RNA that is identified comprise copies which are amplified by the PCR method; preliminarily classifying at least one microorganism by identifying the microbial DNA/RNA by a melting curve; adding at least one labeled oligonucleotide whereby temperature-dependent hybridization takes place and identifying the DNA/RNA on the basis of a physical property of the resultant DNA/RNA oligonucleotide complex, such as the temperature-dependence of the hybridization.

The invention relates to the field of molecular biology and discloses a kit and a method for extracting microbial DNA. The kit comprises lysis solution, inhibitor removal solution and binding solution, wherein the lysis solution comprises 50 to 200mM of Tris-HCl, 50 to 150mM of EDTA, 0.5 to 3M of NaCl, 0.5 to 2 percent of CTAB, 0.5 to 2 percent of PVP and 0.5 to 2 percent of SDS; the inhibitor removal solution comprises 100 to 300mM potassium acetate, sodium acetate or ammonium acetate and 50 to 200mM aluminum sulfate, ammonium sulfate or aluminum ammonium sulfate; and the binding solution comprises 3 to 6M guanidine hydrochloride, 10 to 50mM Tris-HCl and 5 to 50 percent isopropanol. The kit and the method for extracting the microbial DNA have the advantages of high purity, high universality, high extraction speed, direct use for a downstream experiment, and application to extracting DNA from a microbe-containing sample.

The invention relates to a pathogenic microorganism DNA detection chip and the chip comprises a carrier and a nucleic acid probe on the carrier. The nucleic acid probe is provided with a target detection probe used for detecting target gene of pathogenic microorganism. The pathogenic microorganism comprises but not limits to vibrio cholerae, pathogenic escherichia coli, campylobacter jejuni, Yersinia enterocolitica, parahemolytic vibrio, salmonella, and shigella and Listeria monocytogenes. The invention also relates to a preparation method of the pathogenic microorganism DNA detection chip and provides applications of the pathogenic microorganism DNA detection chip in detection of pathogenic microorganism. Also a pathogenic microorganism DNA detection chip kit is provided.

The present invention relates to molecular biology, microbiology, and medicine and provides the method for detection of Mycobacterium tuberculosis complex with simultaneous evaluation of sensitivity of the strains to rifampicin and isoniazid in clinical sample on differentiating biochip. The method is based on two-stage multiplex PCR to obtain fluorescent DNA fragments followed by hybridization of these fragments on microarray containing the set of specific discriminating oligonucleotides. The determination of the resistance of Mycobacterium tuberculosis to rifampicin and isoniazid is carried out by evaluation of point nucleotide substitutions in DNA of microorganism. The present invention allows conduct analysis directly in clinical sample, to evaluate a number of mutations simultaneously, to decrease the cost price of analysis, and to reduce the time of its conducting. The present invention also relates to set of primers, biochip, and set of oligonucleotide probes used in realization of the method.

The invention which belongs to the field of ship ballast water processing equipment especially relates to a method and an apparatus for processing ship ballast water through filtration, ultraviolet and ultrasound. The apparatus of the invention comprises a water pump, a pressure transducer, a flowmeter, a filter, a comprehensive processing unit of ultraviolet and ultrasound, a control system, an electromagnetic valve and a pipeline. Water enters the filter through the water pump, then enters the comprehensive processing unit of ultraviolet and ultrasound to be subjected to biological inactivation, and is finally discharged. The synergetic disinfection of the ultraviolet-ultrasound unit allows the disinfection effect to be good, ultrasonic waves destroy protective layers of microbes, the ultraviolet radiation destroys DNA and RNA of the microbes, and the ultrasonic waves have certain cleaning effects on ultraviolet lamps. The apparatus of the invention, which can effectively kill the microbes in the ship ballast water, has the advantages of low energy consumption, high reliability, simple operation and maintenance, realization of one-key operation through an automatic control unit, and flexible selection of an installation platform according to practical cases.

The invention relates to a simple extraction for the DNAs of microbes in a river environment sample. The method disclosed by the invention comprises: dividing the river environment sample into a water sample and a bottom sediment sample; and extracting the DNAs of the microbes in the water sample and bottom sediment sample respectively. The method can also be used for extracting any environment sample alone. In the invention, by combining a liquid nitrogen repeated freezing and thawing process, a sodium dodecyl sulfate (SDS) process, a lysozyme process and the like, the microbial cells in theenvironment sample can be fully lysed to fully release DNA; and analysis on bacterial diversity by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technique indicates theextraction method can effectively reflect the integrity and diversity of microbes in the river environment sample.

The present invention relates generally to a method for detecting, enumerating and/or identifying microorganisms in a sample. More particularly, the present invention provides a method for determining total microbial content in a sample by detecting the presence of nucleotide sequences associated with all or part of 16S rDNA or its corresponding 16S rRNA or its homologue, functional equivalent or derivative. The nucleotide sequences of the present invention may be used as an indicator of any microorganism and, hence, represents a universal target sequence which is indicative of total microbial content in a sample. The universal target sequence may also be varied to render same genus or species specific or the universal target used to trap microbial DNA or RNA which may be subsequently analyzed by sequence analysis or genetic probe technology. The universal target sequence is useful inter alia to design as universal primers and probes to amplify any microbial-derived genomic sequence, as a means to detect and enumerate total microorganisms and to identify microorganisms in a sample at the genus or species level. Such uses enable improved methods of enviroprotection, bioremediation, medical diagnosis and industrial microbiology. The present invention further relates to the universal target sequence in isolated form and/or primers or probes capable of hybridizing to same and kits for the detection of total microbial content in a sample.

The invention provides an extraction method of total DNA of metagenome of fish enteric microorganisms and a kit. The extraction method comprises the following steps: (1) fish sample sampling and fixing treatment of microorganisms on the surface of the fish; (2) acquisition of enteric microorganisms and collection of thalli; (3) embedding and cracking of cells; (4) extraction of DNA; and (5) DNA preservation. The kit comprises the following main components: a solution A which is a mixed solution of Tris and EDTA; a solution B which is a mixed solution of Tris, EDTA, NaCL, PVP, guanidinium isothiocyanate, Triton-X100, PMSF, and beta-mercaptoethanol; a solution C which is a mixed solution of Tris, EDTA, NaCl and DTT; a solution D which is a mixed solution of proteinaseK, a lysozyme, Tris, NaCl and SDS; a solution E which is a mixed solution of Tris saturated phenol and chloroform; F which is isopropanol; and G which is an ethanol aqueous solution. The DNA fragment obtained by the invention is high in purity, and the content of DNA of a host and the microorganisms on the surface of the fish is small.

The invention belongs to the fields of soil microorganisms, biochemistry and molecular biology and relates to a method for separation and purification of a large-fragment DNA from soil. The method is used for indirect separation and purification of a large-fragment DNA having molecular weight more than 30kb from various types of soil, wherein the large-fragment DNA is used for construction of a metagenomic library, or is used for separation of soil microbial gene clusters. The method solves the problem that separation of microbial cells from soil and preparation of large-fragment soil DNA having high purity and satisfying various biological study demands are realized difficultly by the existing indirect method, and is an efficient method for extraction of soil microorganism DNA having a large fragment and high purity.

This invention relates to an extracting method of soil microorganism DNA. The stated method includes steps as following: cell disruption of soil microorganism, obtaining crude DNA solution, obtaining soil microorganism DNA; the stated method of this invention is using PVPP, protease K etc for valid combination. After organic solvent such as PEG20000, isopropyl alcohol and so on are precipitated, use spectrophotometer to determine value of OD260/OD230, OD260/OD280 close to standard value. It can be directly used for molecule operation, possessing great application perspective.

The invention discloses a DNA extracting method for evaluating the community diversity of the intestinal microorganisms of animals. Before cell disruption, a sample is pretreated, and DNA and pollutants outside the cells are removed, thereby avoiding the problems of difficult removal of the pollutants and low recovery rate of the DNA in a purification step. In the extracting process of the invention, phenol and chloroform are not used, and damage to the health of experiment personnel is reduced; and the obtained DNA is integral and has high yield, and the molecular fragment is larger than 10kb. The OD260/OD230 and OD260/OD280 of the DNA of the extracted intestinal microorganisms are close to standard values and can be directly applied to molecule operation to evaluate the community diversity of the intestinal microorganisms of animals.

The invention relates to a kit and method for extracting a microbial genome DNA by a magnetic bead method. The kit comprises a microbial preservation buffer solution, a bacteriolytic agent solution I, proteinase K, silica gel magnetic beads, isopropanol and an ethanol water solution, and further comprises a bacteriolytic agent II, a lysis adsorption solution and a reinforced scrubbing solution. The bacteriolytic agent II comprises MES, EDTA, Triton X-100, lysozyme and a lysozyme enhancer. The lysis adsorption solution comprises Tris-HCl, sodium chloride, EDTA, TrionX-100 and a nucleic acid separation and absorption promoter; and the reinforced scrubbing solution comprises Tris-HCl, lithium chloride and ethanol. By adopting the kit provided by the invention, cell walls of gram-positive bacteria and various fungi can be completely destroyed to form protoplasts, DNA of the protoplast cells can be rapidly released under the action of the lysis adsorption solution, the DNA is efficiently adsorbed on the silica gel magnetic beads, and after the DNA is washed by the scrubbing solution and eluted by an elution solution, microbial DNA with high concentration and purity can be obtained.

The invention provides a method for purifying microorganism DNA in forest soil, which comprises the following steps: (1) obtaining the rough DNA of a microorganism by a conventional method; (2) dissolving the rough DNA by extracted buffer solution; after the rough DNA is subjected to water bath and isovolumetric chloroform-isoamyl alcohol solution extraction, adding isopropanol which is 0.6 time of the volume of supernatant into the supernatant to precipitate DNA sediments; washing by 70 percent of pre-cooled alcohol and dissolving in the TE buffer solution; and (3) further purifying the initially purified DNA by a silica gel column. The method can remove most of impurities in a rough product of the extracted soil microorganism DNA and obtain the DNA with high purity and can be directly used for the conventional polymerase chain reaction (PCR) which is quite sensitive for an inhabiting matter. The invention has simple operation, low cost, less time consumption and wide application range and can be used for purifying the soil DNA in various types after other prior soil direct extraction methods.

The invention relates to a method for rapidly separating bacteria and a particular primer thereby, in particular to a method for rapidly separating anaerobic denitrifying bacteria and a particular primer thereby, solving the problems that the prior anaerobic denitrifying bacteria have long separation period and great workload and are difficult to obtain bacterial strains and pure bacterial strains with the destination functions. The method comprises the following steps: 1, extracting microorganism DNA from a sample for PCR augmentation and separation culture and prescreening the anaerobic denitrifying bacteria; 2, preparing particular culture media to be purified until a colony grows up; 3, carrying out the enrichment culture of the colony for the PCR augmentation and re-screening the anaerobic denitrifying bacteria; 4, repeating the separation culture, the purification culture and the enrichment culture, identifying and carrying out the PCR augmentation for removing repetitive bacterial strains and then, repeating the separation culture and the purification culture to obtain the pure bacterial strains. An upstream primer is SEQ ID of NO.1, and a downstream primer is SEQ ID NO.2.The invention shortens the separation period by 40 percent to 50 percent, has little workload and is easy to obtain destination bacterial strains and pure bacterial strains.