171 results about "TE buffer" patented technology

The invention discloses a human feces DNA extraction method. The prior art is complicated and time-consuming; and the trace DNAs extracted by the previous sampling method is too little to perform molecular diagnosis. The extraction method uses selective sample collection PSC equipment for feces samples and adopts a low temperature preservation method. The extraction method comprises the following steps: picking a sample by using a sterilized gun head or sterilized blade and directly placing the picked sample on ice; and adding a feces lysis solution, fully cracking and evenly mixing, adding bovine serum albumin, centrifuging to remove the supernatant, adding protease K to perform a digestion reaction at 58 DEG C, then adding a protein precipitation solution to precipitate the protein, centrifuging to obtain the supernatant, adding a silica gel membrane binding solution to fully bind DNAs, centrifuging to obtain precipitates, adding a silica gel membrane washing solution to wash twice, centrifuging, and adding a TE (Tris + EDTA) buffer solution to finally obtain a preservation solution with DNAs. The human feces DNA extraction method has the advantages of high repeatability and stability, simpleness and short time.

The invention relates to a preparing method and application of an amniotic membrane, in particular to a preparing method and application of an acellular amniotic membrane used for repairing skin wound difficult to heal. The preparing method specifically comprises the steps of isolating, cleaning and sterilizing an amniotic membrane tissue; arranging and drying the amniotic membrane tissue on a nitrocellulose membrane to manufacture an amniotic membrane paster; vibrating and digesting the amniotic membrane paster in mixed digestive fluid of 0.25%-0.5% of pancreatin and 0.2-0.5g/L EDTA.4Na for 10-30 minutes at a temperature of 37 DEG C; rinsing the amniotic membrane paster in TE buffer solution; vibrating overnight to fully remove amniotic membrane cells in TE-TritonX-100 solution; washing the amniotic membrane paster for three times in TE solution; vibrating and degrading the amniotic membrane paster for 2-4 hours in nucleic acid degrading solution at a temperature of 37 DEG C; washing the amniotic membrane paster for three times in the TE solution. The prepared acellular amniotic membrane paster can be stored for a long time after being refrigerated, dried, packed and sterilized by cobalt 60 irradiation and is a tissue engineering material which can be taken and used when needed. The acellular amniotic membrane paster can be easily separated from the nitrocellulose membrane to be used for repairing the skin wound after rewatered.

The invention relates to an extraction method of total DNA of microorganisms from a soil sample. The extraction method particularly includes following steps: (1) adding a DNA extraction buffer solution to the soil sample and performing repetitive freeze-thawing in liquid nitrogen and hot water bath; (2) adding a protease K, performing a water bath treatment process at 37 DEG C for 30 min; (3) adding a sodium dodecyl solution, performing a water bath treatment process at 65 DEG C for 30 min; (4) performing centrifugation to remove precipitations and collect a supernate; (5) performing extraction with a mixed solution containing phenol, chloroform and isoamylol and performing centrifugation to collect a supernate; (6) performing extraction with a mixed solution containing chloroform and isoamylol and performing centrifugation to collect a supernate; (7) adding isopropanol to the supernate in the step (6) to performing precipitation and collecting a precipitation after centrifugation; (8) adding ethanol for washing the precipitation and drying the precipitation at the room temperature; (9) adding a TE buffer solution for dissolving the precipitation to obtain a DNA crude extraction; and (10) purifying the DNA crude extraction through an adsorption column, packaging the purified DNA crude extraction and storing the purified DNA crude extraction at -20 DEG C. The extraction method is especially effective when being used in samples from deep soil and dry sand which are less in biomass and in a clayed soil sample. The DNA extraction can be directly used in subsequent molecular biological detection.

Apparatus and method for measuring the fluorescence of nanodrop liquid samples is described in which the sample is held by surface tension between two anvil surfaces. Each anvil surface has an embedded optical fiber with its end finished flush with the surface in the containment area wetted by the sample with the fiber in line. Sample excitation is provided from the side of the sample remote from the containment area. By selection of the fiber transmission numeric aperture the impact of exciting and ambient light on the measurement is minimized. A method of virtual filtering is taught in which any ambient or exciting light that does impinge on the measuring sensor is corrected by subtracting a scaled representation of the source from the measurement. The method and apparatus is capable of detecting 1 femptomole of sodium fluorescein in 1 microliter of TE buffer.

A method for detecting the signals of gene chip whose oligonucleotide has a length more than 50 bases includes such steps as PCR amplification of target gene, detecting, boiling positive product for modifying it, cooling on ice, wetting the gene chip by phosphoric acid solution, prehybridizing in prehybridizing liquid, adding said modified DNA, hybridizing, water with different lotions, dyeing, rinsing, and detecting signal under UV light or fluorescent microscope.

The invention discloses a method for extracting sedum spectabile genomic DNA, comprising the following steps: taking powdered sedum spectabile, and adding preheated 2% CTAB extracting solution to dissolve; carrying out warm bath at 65 DEG C for 45 minutes, adding hloroform and isoamyl alcohol mixture of same volume, mixing, and centrifuging for two times at high speed at room temperature; taking a supernatant, adding ice-cold isopropanol which is 2/3 the volume of the supernatant, mixing, standing at low temperature for 15-30 minutes, and settling DNA; centrifuging at high speed at low temperature, discarding the supernatant, recovering the precipitate, washing for two times by 70% ethanol, airing, adding TE buffering solution, adding RNase, digesting for 30 minutes at 37 DEG C, and preserving the obtained DNA sample at low temperature. According to the method, the added RNase which is an enzyme can remove the RNA thoroughly and effectively so that the extracted DNA has higher purity and the interference of the RNA to the subsequent experiment is avoided.

The invention relates to soil microbiology, in particular to a method of extracting the total DNA of the soil sample, which can improve the quality of DNA. The method includes the following steps: adding DNA extracting buffer to the soil sample, and then adding protease K; freeze thawing under -65 to 65 DEG C and centrifuging; discarding the precipitate; collecting the supernatant liquid; extracting and centrifuging by the same volume of the solution of phenol, chloroform and isoamylol, reserving the water phase; extracting one more time by the solution of chloroform and isoamylol; adding sodium acetate and cold isopropanol, and then centrifuging and collecting the DNA precipitate; washing by ethanol and drying in room temperature; adding TE buffer to dissolve, and gaining the DNA coarse extract; adding DNA extracting buffer in the coarse extract; and then adding the same volume of the solution of phenol, chloroform and isoamylol; repeating the preceding steps such as precipitating, washing and dissolving; finally, gaining the DNA of better quality. The method adopted by the invention can effectively remove the impurity in the DNA, which evidently increases the purity and quality of DNA.

The invention relates to a kit and a method for detecting Macrobrachium rosenbergii Nodavirus. The kit comprises a sampling tube, a rinsing tube filled with distilled water, a nucleic acid denaturation tube filled with TE buffer solution, an amplification test tube filled with amplification reaction liquid and dyes, a negative control tube, a positive control tube, an FTA membrane, quick drying liquid for shortening time of preparing a sample to be detected, namely nucleic acid, and the like. The kit combines the advantages of isothermal amplication and quick dye detection, does not need precious instruments in the detection process, and develops by adopting a dye filling method to improve the detection reliability. The kit has the advantages of low detection cost, convenient use and safety for human bodies and environment, and can replace the conventional related detection methods. The kit can be used for production field in the open field, and has important promotion and application value of enhancing the monitoring of the Macrobrachium rosenbergii Nodavirus and preventing the large-scale outbreak of the Macrobrachium rosenbergii Nodavirus.

The invention discloses a method for extracting a high-quality cell nucleus DNA (Deoxyribonucleic Acid) of a plant rich in polysaccharide and polyphenol, relating to the field of plant functional genomics. The method comprises the following steps of: firstly, grinding a plant material by using liquid nitrogen; secondly, resuspending by using an HB (Haemoglobin) extraction buffer solution, mixing uniformly through volution and filtering; thirdly, centrifuging, washing precipitation with an extraction buffer solution 1 three times and resuspending; fourthly, carrying out water bath by using an extraction buffer solution 2 and RnaseH at the temperature of 65DEG C for 30 minutes; fifthly, extracting once by using chloroform-isoamylol and precipitating by using isopropanol; and sixthly, washing precipitation by using 75 percent pre-cooled ethanol 1-2 times and dissolving by using a TE (Tris-Ethylene Diamine Tetraacetic Acid) buffer solution. According to the pretreatment step in the methoddisclosed by the invention, DNAs of cell organelles such as mitochondria and chloroplast can be effectively removed; the inference on the nucleic acid extraction step caused by a large mount of secondary metabolites such as polysaccharide and polyphenol existing in the plant material is avoided; and the method is suitable for healthy tissues of wide plant species or frozen tissues with more complete cell structures and is especially suitable for high throughout sequencing in the subsequent researches.

The invention discloses a high-purity plasmid extraction kit. The high-purity plasmid extraction kit comprises an S I buffer solution, an S II buffer solution, an S III buffer solution, a PS cleaningsolution, an elution buffer solution, a TE buffer solution and a novel centrifugal column, wherein the S I buffer solution comprises Tris-HCl of 1 M, EDTA of 0.5 M and glucose of 1 M, and the pH valueis 8.0; the S II buffer solution comprises NaOH of 2 M and 10% SDS; the buffer solution S III comprises potassium acetate of 3 M, guanidine hydrochloride of 2 M and acetic acid of 2 M, and the pH value is 3.6-4.2; the PS cleaning solution is an isopropyl alcohol solution containing 5-10% of TritonX-114; and the elution buffer solution is 75% ethanol. The invention further provides a method for extracting a large number of plasmids by using the kit. According to the method, the novel centrifugal column is combined with a classic strong base-SDS bacterial cell cracking method, so that a plasmidDNA sample is centrifugally combined to a purification column, plasmid DNA can be fully eluted under a certain condition, rapid purification of plasmids is realized, phenol chloroform extraction is not needed in the whole process, high-purity plasmid DNA can be obtained, and the use of eukaryotic cell transfection can be met.

The invention discloses a PCR diagnostic kit for porcine infectious pleuropneumonia, which comprises 400mu L of proteinase K with the concentration of 20mg/mL, 1000mu L of cracking solution, 1500mu L of TE buffer solution, 250mu L of PCR enzyme, 170mu L of ultra-pure water, 50mu L of MarkerDL2000, 40mu L of primer P1 and primer P2 which are mixed in the same volume and have the same concentration of 20mu M, 20mu L of negative control and 20mu L of positive control. By optimizing the PCR reaction conditions, the invention develops a PCR kit for detecting the actinobacillus of the porcine infectious pleuropneumonia; and by comparing the PCR detection result with the negative control and the positive control, detection conclusions can be obtained, thereby achieving the aim of quickly detecting whether a sample has porcine infectious pleuropneumonia or not. The invention has accurate detection result, quick and sensitive detection process, simple detection mode and good using effect.

The invention belongs to the technical field of molecular biology, and discloses an extraction method of microorganism genome DNA. The extraction method comprises the following steps: (1) collecting sample thallus, adding a STES buffer solution, re-suspending, adding glass beads, and dissolving by virtue of a TE buffer solution; (2) adding an equal volume of phenol/chloroform solution, oscillating, mixing, and primarily centrifuging; (3) adding an equal volume of chloroform/isoamylol solution into an upper layer of water after the primary centrifuging, mixing, and secondarily centrifuging; (4) adding an equal volume of isopropanol or twice volume of anhydrous ethanol into an upper layer of water after the secondary centrifuging, precipitating under the room temperature, tertiarily centrifuging, and collecting precipitates; (5) washing the precipitates by utilizing ethanol, re-centrifuging to obtain DNA precipitates, dissolving the DNA precipitates by utilizing sterile water, carrying out the magnetic-ball passivation, to obtain extracted microorganism genome DNA. The genome DNA obtained by virtue of the method has advantages of high concentration, single banding and no pollution, and satisfies the three-generation sequencing requirement.

The invention provides a saliva preservation solution, and a preparation method and application thereof. The saliva preservation solution comprises a DNA stabilizer including disodium edentate, tartaric acid and at least one selected from a group consisting of nitrilotriacetic acid and dihydroxyethylglycine; a microbial inhibitor including penicillin, streptomycin, geneticin and at least one selected from a group consisting of potassium sorbate, sodium benzoate, ethylparaben sodium, ketoconazole and puromycin; and a buffer solution including a TE buffer solution and at least one selected from a group consisting of a PBS buffer solution and a sodium chloride glucose buffer solution. According to the invention, through preparation of the saliva preservation solution, the morphology of cells in a saliva swab is maintained and prevented from disruption, and the growth of microorganisms in the saliva swab and the saliva preservation solution is inhibited.

The fast extraction method of adnascent microbe community total DNA of sponge includes cutting sponge sample into small blocks, suspending on TE buffering liquid and set on ice for direct grinding to cracking sponge tissue, adding lysozyme, sodium dodecyl sulfonate and proteinase K into the cracked sponge, centrifuging and adding Tris-saturated phenol to extract, extraction with Tris-saturated phenol, chloroform and isopentanol solution for several times, adding NaCl and isopropanol to obtain DNA precipitate, washing and dissolving in TE buffering solution, slaking with RNA enzyme, and final agarose gel electrophoresis analysis to detect extracted DNA sample. The said method can obtain total DNA sample of adnascent microbe community in large segment of sponge for the need of adnascent microbe community structure and diversity analysis based on nucleic acid.

The invention discloses a general nucleic acid isothermal amplification detection kit for pathogenic vibrios of mariculture animals. The detection kit comprises a grinding liquid tube into which grinding liquid is filled, a nucleic acid extracting solution tube A into which solution of sodium acetate is filled, a nucleic acid extracting solution tube B into which absolute ethanol is filled, a nucleic acid extracting solution tube C into which 70 mass percent ethanol solution is filled, a tris-hydrogen chloride ethylene diamine tetraacetic acid (TE) buffer solution tube into which TE buffer solution is filled, a uracil-DNA-glycosylase (UNG) tube into which uracil-DNA-glycosylase is filled, a loop-mediated isothermal amplification (LAMP) reaction liquid tube into which LAMP reaction liquid is filled, a bacillus stearothermophilus deoxyribonucleic acid (Bst DNA) polymerase tube into which Bst DNA polymerase is filled, a color-developing agent tube into which a nucleic acid dye SYBR Green I is filled, a positive control nucleic acid tube into which positive DNA of the vibrios is filled and a negative control tube into which sterilized double distilled water is filled. The invention also discloses a method for detecting the pathogenic vibrios of the mariculture animals by utilizing the detection kit.

The invention provides a concentration-gradient fluorescent calibration piece for a laser microarray chip scanner. The concentration-gradient fluorescent calibration piece comprises a glass substrateand an organic fluorescence mixed liquor, wherein the organic fluorescence mixed liquor is uniformly distributed on the glass substrate in a lattice manner and comprises an Alexa Fluor 488 fluorescentdye, a Cy3-dCTP fluorescent dye and a diluent, the diluent is a phosphate buffer liquor, a TE buffer liquor or an aqueous liquor, and the surface of the glass substrate is chemically modified or coated with a polymer piece. The lattice of the organic fluorescence mixed liquor on the glass substrate comprises 1-8 branch lattices, each branch lattice is arranged on the glass substrate in a uniformdistribution manner, organic fluorescence mixed liquor droplets of each branch lattice are arranged in A rows* B lines, A ranges from 20 to 40, and B ranges from 5 to 20. The concentration-gradient fluorescent calibration piece has the beneficial effects that the concentration-gradient fluorescent calibration piece is simple to produce, short in production period and low in cost and has outstanding detection effect.

The invention discloses a method of extracting high molecular genome from the bacteria. The method contains the steps of: breaking cell wall using lysozyme +protease +SDS or protease +SDS, extracting in TE buffer (removing proteins and polysaccharides), and preciping, washing, drying and dissolving. The invention begins with the factors influencing extracted DNA fragment length and obtains high molecular DNA fragment longer than fragments extracted by conventional method. The genomic DNA extracted in the method has more than 40 kb with OD260/OD280 between 1.8 and 2.0 detecting by absorbance. The method has the advantages of good repeatability and simple operation and can meet the need of the above molecular operation.

The invention discloses a method for extracting bacterium genomic DNA by using magnetic nanoparticles, belonging to the technical field of nanomaterials and molecular biology. The method comprises the steps of: preparing a magnetic nanoparticle aggregate, preparing a bacterium lysing solution, preparing and adding an absorption buffer solution, absorbing with a magnet, rinsing with 70 percent alcohol, and eluting with a TE buffer solution to obtain a bacterium genomic DNA solution. Compared with the traditional phenol/chloroform DNA extracting method, the method for extracting the bacterium genomic DNA by using magnetic nanoparticles has the advantages of short experiment time, simpleness in operation, high extraction efficiency, rapidness, convenience, sensitivity, high yield and the like; and the extracted bacterium genomic DNA can be completely used for subsequent experiments, such as DNA hybridization, PCR (Polymerase Chain Reaction), and the like, and provides the basis for the subsequent research and analysis.

The invention relates to an on-site rapid high-sensitivity detection kit of prawn infectivity muscle necrosis virus and a detection method thereof. The detection kit comprises a sampling tube, a rinsing tube filled with distilled water, a nucleate degeneration tube filled with a TE buffer solution, an amplification detection tube filled with amplification reaction liquid and nucleic acid dye, a negative control tube, a positive control tube, an FTA membrane, rapid drying liquid, and the like. Compared with the RT-PCR detection method, the detection method in the invention has the advantages of higher specificity, sensitivity and convenience, low cost, convenient use, more accurate and rapid detection and extremely sensitivity, is safe to both human and the environment and can substitute the traditional relevant detection method. The invention can be used in both the indoor laboratory and outdoor production field, has great significance for strengthening the epidemiological monitoring of prawn infectivity muscle necrosis virus, prawn cultivation waterbody monitoring and disease prevention and control and has great promotion and application value.

The invention belongs to the technical field of environmental biology, and specifically relates to a PCR template simple preparation method and a PCR amplification method that can be used in large-scale fungi strain identification. With the methods, fungi DNA extraction is not required. According to the fungi DNA simple preparation method, a PCR tube containing a small amount of mycelium and a small amount of a TE buffering solution is heated and broken by using a PCR amplification apparatus, such that a small amount of fungi DNA is obtained; combined with a modified PCR amplification system, a fungi DNA fragment required by the fungi identification is obtained. The method is advantaged in less steps, low cost, simple operation, good amplification effect, and environment-friendliness. The method is suitable for large-scale fungi molecular biological identifications.

The invention belongs to the technical field of molecular biology, and discloses an extraction method of strawberry genome DNA. The extraction method comprises following steps: liquid nitrogen is used for smashing samples into powder; CTAB extracting solution is added; an obtained mixture is subjected to water bath at 55 to 65 DEG C and then is cooled to room temperature; potassium acetate is added, and ice bath is carried out; a chloroform/isoamyl alcohol mixed liquid of a same volume is added; an obtained mixed material is mixed uniformly, and is subjected to centrifugation; the chloroform/isoamyl alcohol mixed liquid of a same volume is added into an obtained supernate A; an obtained product is mixed uniformly, is allowed to stand, and is subjected to centrifugation so as to obtain a supernate B; isopropyl alcohol is added into the supernate B, an obtained mixed solution is subjected to centrifugation so as to obtain a precipitate; a supernate C obtained via centrifugation is removed; TE buffer containing RNase is subjected to water bath at 37 DEG C; the chloroform/isoamyl alcohol mixed liquid of a same volume is added, an obtained mixed liquid is subjected to uniform mixing, is allowed to stand, and is subjected to centrifugation so as to obtain a supernate D; sodium acetate and absolute ethyl alcohol are added into the supernate D; an obtained mixed product is subjected to centrifugation so as to remove an obtained supernate E; TE buffer is added into an obtain precipitate product so as to dissolve the precipitate product, and a strawberry genome DNA solution is obtained. According to the extraction method, characteristics of CTAB and SDS extraction methods are combined, and high quality genome DNA of strawberry which is abundant in glucose and polyphenols is obtained.

The invention discloses an extraction method of the genome DNA of isolated soy proteins, comprising the following steps of: firstly sufficiently mixing isolated soy proteins and a TE buffer solution to prepare a TE mixed solution; then adding a guanidinium isothiocyanate lysis solution with volume twice larger than that of the TE mixed solution, sufficiently uniformly mixing, and lysing at room temperature; adding isometric phenol and chloroform/isoamylol to extract proteins; centrifugalizing, and then adding isometric chloroform/isoamylol to a supernate to extract the proteins; centrifugalizing, and then adding isometric chloroform to the supernate to extract the proteins; centrifugalizing, and precipitating the supernate by using isopropanol; centrifugalizing, and washing by using ethanol for precipitation; airing at the room temperature; and dissolving in the TE buffer solution so as to obtain the TE solution of the genome DNA. According to the extraction method, the genome DNA which has high quality and is suitable for PCR (Polymerase Chain Reaction) detection is extracted from the isolated soy proteins, the concentration of the genome DNA is 1558 micrograms/ml, and the value of OD260/OD280 is 1.666; and in addition, the invention has the advantages of easy operation and short consuming time and is beneficial to fast detection.

The invention relates to a method to separate isomeric protein form gene recombinant expression protein. The method of the invention is that: the collected wet bacterium is made into bacterium soliquoid in a cleaning buffer liquid, and shattered and decentered by ultrasonic, and the clear liquid is abandoned, the precipitation is cleaned by TE buffer liquid containing urea and TritonX-100, then protein endosome is acquired, and the endosomes is fully dissolved by a buffer liquid containing guanidine hydrochloride or urea; the dissolving liquid is diluted and renatured by the buffer liquid containing Tris-HCl, arginine and EDTA, then stand for 24-48h, and becomes a CNTF renatured protein liquid; after being ultrafiltered and concentrated, the renatured protein is transferred into the Tris buffer liquid through gel filtration chromatography to collect protein peak; finally the CNTF renatured protein liquid with transferred liquid is sent to Q sephrose-Fast Flow anion exchange column, to remove unrenatured CNTF proteins and other proteins.

The invention relates to a kit capable of rapidly detecting the cryptosporidium in food. The kit mainly solves the technical problems of the absence of a detecting method at both home and abroad and too long detection time. The kit capable of rapidly detecting the cryptosporidium in food consists of the following agents: an agent A: a fixed-DNA FTA card consisting of an FTA card, an FTA purifying reagent and an FTA cleaning reagent (TE buffer:0.01mol/L Tris, pH 8.0; 0.1 mmol/L EDTA); an agent B: PCR reaction liquor containing two specific primer pairs of CryoutL and CryoutR which respectively have the sequences of 5'-TTCTAGAGCTAATACATGCG-3' and 5'-CCCACTT CTTCGAAGCAGGA-3'; an agent C: PCR reaction liquor containing two specific primer pairs of CryinL and CryinR which respectively have the sequences of 5'-GGAAGGGTTGTATTTATTAGATAAAG-3' and 5'- CTCATAAGGTGCTGAAGGAGTA-3'. The method is used to detect the cryptosporidium in a sample.