95 results about "Desoxyribonucleic acid" patented technology

The invention discloses applications of a CRISPR/Cpf1 system in gene editing. CrRNA in the CRISPR/Cpf1 system is modified crRNA; the modification manner is desoxyribonucleic acid modification; the desoxyribonucleic acid modification method comprises the step of increasing 1-3 desoxyribonucleic acid at the 5' terminal and/or increasing 1-3 desoxyribonucleic acid at the 3' terminal of crRNA; and the desoxyribonucleic acid is deoxythymidine acid. The experiment proves that the crRNA formed through chemical synthesis and having the modification can cause the mutation of an hAAVS1 gene and a THUMPD3-AS1 gene when used for the AsCRISPR/Cpf1 system or the FnCRISPR/Cpf1 system, namely, the modified crRNA has certain gene editing capacity when used for the AsCRISPR/Cpf1 system or the FnCRISPR/Cpf1 system, and therefore, the modified crRNA has a significant application value in gene editing utilizing the CRISPR/Cpf1 system.

The invention discloses a plasma free nucleic acid extraction kit and application thereof, wherein the kit comprises (1) magnetic bead pyrolysis combination solution; (2) a first cleaning solution; (3) a second cleaning solution; (4) an elution solution. By using the plasma free nucleic acid extraction kit provided by the invention, the free desoxyribonucleic acid in the plasma can be fast extracted; in addition, the use is fast, simple and convenient; the centrifugation is not needed; once mass extraction only needs half an hour; the obtained nucleic acid has high quality; when the obtained nucleic acid is used for subsequent sequencing and detection, the result is accurate and reliable; the complete application to the existing noninvasive prenatal genetic testing technology can be realized; the important significance is realized in aspects of scientific study and clinic detection.

The invention relates to a DNA (Desoxyribonucleic Acid) ligand for specific binding with EPO (erythropoietin), a preparation method thereof and application thereof, specifically provides a DNA ligand capable of specifically binding with the EPO and a method for screening the ligand by an improved SELEX (Systematic Evolution of Ligands by Exponential Enrichment) technology, and provides EPO detecting elements with better detecting stability, higher sensitivity, convenience and quickness.

The invention discloses a CRISPR-Cas based technology for normal-temperature/isothermal rapid detection of ribonucleic acid and a detection method. The invention aims to perform in-vitro ribonucleic acid amplification on multiple disclosed gentically engineered enzymes and chemical components and to realize amplification and detection of specific ribonucleic acid (RNA) or desoxyribonucleic acid (DNA) quickly in one step or two steps in normal-temperature/isothermal condition so that the detection of each nucleic acid is faster and more convenient. The method comprises the following steps: (1)performing nucleic acid extraction to obtain the RNA, single-stranded DNA or double-stranded DNA of a to-be-detected sample; (2) conducting a reaction with to-be-detected nucleic acid by using the combined enzyme of reverse transcriptase, transcriptase, ribonuclease H and CRISPR-associated protein 13 as well as a nucleic acid fluorescent probe in an isothermal condition, wherein if the to-be-detected nucleic acid is DNA, a pre-denaturation step is added before the reaction; (3) detecting the fluorescence signal to judge whether target nucleic acid exists in the to-be-detected sample.

The invention discloses an array micro polymerase chain reaction chip, and relates to desoxyribonucleic acid (DNA) amplification technology. The chip comprises a temperature control system, a heating chip and a reaction cavity; the heating chip and the reaction cavity are arranged separately; and during use, after a reaction cavity framework is sleeved on the external circle of an insulating substrate, the reaction cavity framework is pressed down so that the lower surface of the bottom of a reaction tank is tightly jointed with the upper surface of a heating electrode, sample solution is injected into the reaction tank, and then the chip is started. The amplification reaction can be performed by sticking a clean disposable polydimethylsiloxane (PDMS) micro array to the heating chip every time. The reaction chip can microminiaturize polymerase chain reaction (PCR), fulfills the purposes of microminiaturization, lightness, quickness and saving, and can also solve the problems of biocompatibility difficult to be overcome by microminiaturization and pollution of the PCR.

The invention discloses a method for extracting total RNA (Ribonucleic Acid) from woody mangrove plants and special extracting solution thereof. The extracting solution provided in the invention comprises Tris-HCl, EDTA (Ethylene Diamine Tetraacetic Acid), NaCl, SDS (Sodium Dodecyl Sulfonate), DTT (Dithiothreitol), beta-mercaptoethanol and water, wherein the final concentration of the Tris-HCl is 75-125 mmol/L, the final concentration of the EDTA is 400-600 mmol/L, the final concentration of the NaCl is 400-600 mmol/L, the final concentration of the SDS is 1%-3% (in weight percent), the final concentration of the DTT is 3-8 mmol/L, and the final concentration of the beta-mercaptoethanol is 1%-3% (in weight percent); and the pH value is 8-9. The method provided in the invention is used for extracting the total RNA from the woody mangrove plants by using the extracting solution. The RNA extracted by the method can be directly used for synthesizing cDNA (complementary Desoxyribonucleic Acid) and carrying out molecular biology operations, such as library establishment, molecular clone and the like.

The invention relates to a method for extracting desoxyribonucleic acid from formalin fixed and paraffin embedded tissues, and belongs to the technical field of nucleic acid application in biology. The method comprises the following steps of: preparing special lysis solution, adding the lysis solution into paraffin section tissues, boiling at high temperature for 30 minutes, and centrifuging to take supernate; adding absolute ethanol for uniform mixing, and adding the mixed solution into a silicon membrane absorption column for centrifuging; rinsing by using protein-free liquid and salt-free rinsing liquid; and eluting by using elution buffer. In the method, a toxic reagent of dimethylbenzene is not used for dewaxing, and harm to bodies of experimenters is avoided; and precious protease Kis not needed to perform long-time incubation enzymolysis, the operation is simple and quick, the extracted desoxyribonucleic acid in genome is high in quality and stability, and the cost and time can be saved to the greatest degree. The extracted product is subjected to polymerase chain reaction (PCR) detection, long segments with about 750pb can be obtained through amplification, and the work in the aspects such as scientific research, biomedicine and the like is greatly facilitated.

Provided is a fluorescence detection method for detecting desoxyribonucleic acid binding protein. The method utilizes double-strand nucleic acid containing protein binding locus and marked by fluorescein as a probe, the probe is bound with target protein to form a nucleic acid probe/desoxyribonucleic acid binding protein composite which provides protection space for the nucleic acid probe, so that the nucleic acid probe can be prevented from being hydrolyzed by nuclease, the nucleic acid probe can be bound with cationic water-soluble conjugated polymer through a static reaction, the distance between the fluorescein and the polymer is small, and high-efficient fluorescence resonance energy transfer can be generated. Based on the water-soluble conjugated polymer and exonuclease III, the method includes the following steps: a, preparing the double-strand desoxyribonucleic acid fluorescence probe capable of binding protein with specificity; b, binding the double-strand desoxyribonucleic acid fluorescence probe with the desoxyribonucleic acid binding protein; c, enzyme restriction can be conducted on the composite through exonuclease III; and d, adding the water-soluble conjugated polymer into the reaction system for fluorescent detection.

The invention belongs to the technical field of the biological medicine, and particularly relates to a cation-modified konjac glucomannan gellan gum microsphere as well as a preparation method and an application. The cation-modified konjac glucomannan gellan gum microsphere is prepared from a nucleic acid drug and a drug delivery carrier, wherein the nucleic acid drug is ribonucleic acid or desoxyribonucleic acid, and a drug delivery system comprises the components as follows: 20-30 parts of cKGM (cation konjac glucomannan), 1-9 parts of gellan gum and 0.01-6 parts of the nucleic acid drug. According to the microsphere, the requirement for colon-specific drug delivery of the oral microsphere is met, and the microsphere is simple in preparation technology and easy to industrialize and has very broad development and application prospects.

The invention provides a freeze-dried microsphere of a PCR reaction agent. The freeze-dried microsphere comprises an enzyme reaction liquid, desoxyribonucleic acid triphosphoric acid, a primer, a template, a biological buffer solution, glucan, trehalose, bovine serum albumin, gelatin, glycerinum, dimethyl sulfoxide, a surfactant, a defoaming agent, a preservative and the like. By means of the freeze-dried microsphere of the PCR reaction agent and the preparing method thereof, the problem that a PCR agent cannot be preserved or transported at a normal temperature in the prior art is solved; theproblems that the PCR agent can hardly be packaged or stored and a freeze-dried powder agent can hardly be quantified are solved. The freeze-dried product comprises a PCR reaction main agent and is convenient to use and saves time, and the contamination introduced in the experiment operation process and the errors of reducing the addition of reaction components are effectively avoided.

The invention discloses a high-throughput method based on DNA (desoxyribonucleic acid) and RNA (ribonucleic acid) gene mutation detection. The method comprises the steps that (1) desoxyribonucleic acid (DNA) and ribonucleic acid (RNA) are extracted from a biological sample collected from a subject through utilizing a reagent, and a DNA library is constructed based on desoxyribonucleic acid; ribonucleic acid is reversely transcribed into double-stranded cDNA firstly, and then is constructed into an RNA library; (2) a first probe set is mixed with the DNA library, and meanwhile, a second probe set is mixed with the RNA library; (3) a complex of probes and nucleic acid is separated and obtained from mixtures obtained in step (2) by utilizing markers in the first probe set and second probe set; (4) sequencing is performed on target nucleic acid in the complex by utilizing a high-throughput technique. The method is based on NGS methodology and optimizes the NGS methodology, so that the specificity and sensitivity of detection are further improved. In addition, a DNA detection result can also be verified by detecting RNA.

The invention discloses a method for constructing a strand-specific transcriptome library. The method comprises steps as follows: step 1), mRNA (massager ribonucleic acid) fragmentation processing is performed; step 2), inverse transcription is performed: the fragmented mRNA 3' terminal is connected with an R3 joint with the known sequence; an RT primer and the R3 joint are subjected to annealing pairing for inverse transcription, and a first strand of cDNA (complementary desoxyribonucleic acid) is synthesized; step 3), cDNA 3' terminal marking is performed: after RNA is removed, TdT (terminal deoxynucleotidyl transferase) is adopted to add multiple dC basic groups to the cDNA 3' terminal, ddCTP (2',3'-dideoxycytidine-5'-triphosphate) is adopted for end closing, pairing is performed with an F5G primer, and the first strand of cDNA is duplicated under the action of polymerase; step 4), PCR (polymerase chain reaction) amplification is performed; step 5), quality testing is performed. The fragmented mRNA is connected with the joint and then is synthesized, so that the synthetic efficiency and the homogeneity of cDNA are improved; after the first strand of cDNA is synthesized, multiple dC basic groups are added to the cDNA 3' terminal by adopting TdT and ddCTP is adopted for closing, pairing is performed with the F5G primer containing multiple dG basic groups, the first strand of cDNA is duplicated under the action of polymerase, therefore, production of self-linked dimers can be effectively reduced, and the homogeneity of the library is improved.

The invention discloses a biological nucleic acid rust inhibitor as well as a preparation method and an application thereof. The biological nucleic acid rust inhibitor contains a solvent and DNA (desoxyribonucleic acid) dissolved in the solvent, wherein the solvent is water or a solution. Compared with the prior art, the biopolymer DNA steel bar rust inhibitor solves the problem that a traditional rust inhibitor has negative effects on reinforced concrete due to higher consumption, biological toxic effect, complicated synthesis method, short acting time and the like; the biological nucleic acid rust inhibitor has the advantages of a purely biological rust inhibitor as follows: the consumption is lower, pollution is avoided, the acting time is long, production process is simple, corrosion of steel bars can be effectively inhibited and reduced and the like.

The invention relates to fast check paper of Staphylococcus aureus causing food poisoning, in particular to a formula of a fast check reagent of food poisoning caused by the intestinotoxin of toxin-production Staphylococcus aureus and a preparation method thereof. The invention breaks through a traditional sectional check means and utilizes the characteristics of heat-resisting desoxyribonucleic acid and plasma-coagulase, the check diagnosis process is shortened to 8-12 hours, and the identification accuracy is larger than 97 percent. The fast check paper has the characteristics of high speed, high accuracy, high sensitivity, and the like. Compared with a traditional method, the fast check paper has an application value, has an important meaning for guiding clinical diagnosis and treatment and avoiding abusing antibacterial medicines, and has an obvious advantage in the application of rapidly identifying and diagnosing toxin-production Staphylococcus aureus which is a food poisoning pathogenic bacterium.

The invention discloses a MicroRNA detection method. The method includes that through combination of a DNA (deoxyribonucleic acid) probe molecule modified with aggregation induced luminescence compounds (AIE molecules) and to-be-detected MicroRNA and under the action of DNA excision enzyme III, hydrophilic desoxyribonucleic acid in a probe can be hydrolyzed, remained hydrophobe AIE substances aggregate to give out light, the targeted MicroRNA is released to be combined with the probe, the probe is hydrolyzed once more, fluorescent light is enhanced after circulation for multiple times, and the MicroRNA is detected. The invention further discloses the probe molecule and a reagent box used for detecting miR-21 with the method. The method used for detection of the MicroRNA is high in speed and low in detection limit, and the probe is simple in process and low in cost.

The invention relates to a method for extracting free nucleic acid from large-volume cell-free body fluid. The method comprises the following steps: S1, concentrating, specifically, adding a precipitating agent and a buffer solution into the cell-free body fluid, evenly mixing by means of vortex and then placing at the room temperature, centrifuging the cell-free body fluid, and discarding a supernatant; S2, cracking, specifically, adding sterile water into precipitate obtained in the S1, evenly mixing upside down, centrifuging the obtained mixture, and discarding the supernatant; then, addinglysate and protein K, resuspending precipitate, and carrying out incubation at high temperature; S3, purifying, specifically, adding isopropanol into the suspension obtained in the S2, evenly mixingand then enabling the obtained mixture to be combined with an adsorption column or magnetic beads, cleaning the sample with washing liquor for a plurality of times, and finally eluting the product with eluant so as to obtain the high-purity free nucleic acid. The desoxyribonucleic acid (DNA) sample extracted by using the method is high in yield and purity and is applicable to the detection, such as fluorescence quantitative polymerase chain reaction (PCR), with higher requirement for a template, so that the universality of subsequent sample detection is improved.

The invention relates to a multi-degree-of-freedom self-assembly nano-robot and a manufacturing control method thereof. The nano-robot is a quadruped nano-robot formed by self-assembly of micro-nano particles and four desoxyribonucleic acid chains through strong interaction of gold-mercapto bonds or streptavidin and biotin under mutual effects. Round gold electrodes are deposited at the four vertexes of a square on a silicon-based material respectively, and four nanopores are machined in the round gold electrodes at the same time; and under the sizes of the nanopores, each nanopore can captureonly one desoxyribonucleic acid chain under the action of an external electric field. The nanogold electrode is connected with an external voltage source; the electrical property and strength of charge density on the nanopore can be regulated and controlled by regulating and controlling the direction and size of voltage on each nanopore, so that the direction and strength of electroosmotic flow passing through the nanopore are controlled and combined or competitive drive is formed by the electric field force borne by the desoxyribonucleic acid chain and thus the movement speed and direction of the nano robot are controlled.

The invention relates to the field of engineering bacterium saccharomyces cerevisiae and plant genetic engineering, and provides application of an OsBBTI4 (oryza sativa Bowman-Birk trypsin inhibitor 4) gene to reduction of heavy metal cadmium accumulation in oryza sativa. The cDNA (complementary desoxyribonucleic acid) overall length sequence of the gene is shown in SEQ ID NO.1, and the encoded amino acid sequence of the gene is shown in SEQ ID NO.2. OsBBTI4 gene encoded protein OsBBTI4 is related to heavy metal cadmium accumulation of oryza sativa. By means of transgenosis overexpression or RNAi(ribonucleic acid interfere) of the OsBBTI4 gene in oryza sativa, the expression quantity of the OsBBTI4 gene in transgenic oryza sativa strains is regulated and controlled, cadmium content of transgenic oryza sativa seeds can be affected, and a low-cadmium-enrichment transgenic oryza sativa strain is obtained. The OsBBTI4 gene can be applied to genetic engineering breeding of engineering bacteria and oryza sativa for changing in-vivo cadmium content, and genetic engineering breeding of other plants for in-vivo cadmium enrichment.

The invention provides a preparation method of amino magnetic nanoparticles and application thereof in DNA (desoxyribonucleic acid) extraction. The preparation method of the amino magnetic nanoparticles comprises the following steps: (1) preparing Fe3O4 magnetic nanoparticles; (2) preparing sodium bitartrate coated Fe3O4 magnetic nanoparticles; and (3) preparing ethidene diamine modified sodium bitartrate coated Fe3O4 magnetic nanoparticles. The amino magnetic nanoparticles prepared by using the preparation method is free of high temperature or high pressure in the synthesis process, are freeof waste, and thus have the characteristics of being green and environment and simple in process; compared with a conventional commercial kit, the nanoparticles are easy in raw material obtaining, lowin price, simple in synthesis step and easy to operate; and the nanoparticles have amino groups on surfaces, are at positive charge states and are beneficial to combination with DNA with negative charges, in the DNA extraction application, the magnetic nanoparticles have good DNA combination capabilities and high extraction efficiency, extraction steps are simple and convenient, and toxic reagents such as chloroform can be avoided.

The invention discloses a hepatitis c virus adsorbent as well as a preparation method and an application thereof, and belongs to the field of medical and biological materials. The hepatitis c virus adsorbent comprises a carrier and a single strand desoxyribonucleic acid ligand combined with a hepatitis c virus envelope protein and connected to the carrier. The sequence of the ligand is as shown in SEQINNO.2. The hepatitis c virus adsorbent is obtained by obtaining a sensitive carrier surface through a chemical activating reaction and performing covalent linkage of the carrier surface with the artificially synthesized single strand desoxyribonucleic acid. The adsorbent is simple and convenient in synthetic method, short in process line and safe to prepare. The covalent bond is firm to connect and the ligand is stable to combine. The product has the characteristics of strong specificity, high absorption efficiency to hepatitis c virus particles, good regenerability and no toxicity and heat source reaction, and can be used for absorptive treatment of clinically and specifically eliminating hepatitis c virus particles in plasma and can be used for preparing anti-hepatitis c virus medicines.

The invention discloses an aqueous phase preparation method of red light silver sulfide quantum dots and belongs to the cross technical field of material science and photonics. The aqueous phase preparation method concretely comprises the following steps: taking silver nitrate and sodium sulfide as reaction precursor materials and desoxyribonucleic acid as a stabilizer, reacting, precipitating and producing nano crystals in an aqueous liquid, and obtaining silver sulfide quantum dots which can be stably dispersed in the aqueous liquid by virtue of a centrifugal separation method. The method disclosed by the invention adopts water as a solvent, the reaction temperature is mild and controllable, the size of the obtained silver sulfide quantum dot product is 1-10 nanometers, and fluorescence emission is located in a red band, so that the aqueous phase preparation method is applicable to fluorescence microscopy imaging detection of in vitro cells and has broad application prospects in the aspect of biological detection.

The invention provides application of a gene segment of a jasmonic acid-isoleucine hydroxylase encoding gene StCYP94B3-like and a silent vector of the gene segment in increasing the yield of potatoesand relates to the technical field of gene engineering. The cDNA (complementary desoxyribonucleic acid) sequence of the gene segment of StCYP94B3-like is shown in SEQ ID NO.1 in the description. Afterthe RNAi (ribonucleic acid interfere) silent vector is constructed by using the StCYP94B3-like gene segment and transformation of the silent vector into potatoes, the content of jasmonic acid-isoleucine (JA-Ile) can be remarkably increased, the content of jasmonic acid (JA) is not largely affected, and furthermore, the yield of the potatoes can be increased.

The invention relates to a method and a kit for extracting total desoxyribonucleic acid in crude oil. The method comprises the following steps of carrying out pretreatment on the crude oil; cracking a viable cell and/or a dead cell by a positive-negative surfactant method to obtain a supernatant fluid containing the total desoxyribonucleic acid; extracting the supernatant fluid to obtain the total desoxyribonucleic acid by utilizing one of four purification methods of chloroform-alcohol precipitation, phenol/chloroform-alcohol precipitation, a chloroform-purification column and a phenol/chloroform-purification column. According to the method and the kit, aiming at the self property of the crude oil and the characteristic that components are complicated, four methods and kits for extracting the total desoxyribonucleic acid in the crude oil, which are wide in scope of application, are developed; the total desoxyribonucleic acid in the crude oil can be effectively extracted.

The invention provides a traditional Chinese medicinal preparation for conditioning and treating chronic hepatitis B, and belongs to the fields of traditional Chinese medicines and health care. The following raw materials are combined: peach kernel, fingered citron, fructus aurantii, endothelium corneum gigeriae galli, coix seeds, cordate houttuynia, dandelion, honeysuckle, herba portulacae, cape jasmine fruit, dark plum, liquorice root and jujube, so that by virtue of the synthetic effect of the various medicines, the preparation has the functions of promoting blood circulation and removing stasis, tonifying spleen and removing damp, strengthening healthy qi and nourishing liver, and clearing away heat and toxic materials, and therefore, an effect of conditioning and treating the chronic hepatitis B is achieved. Clinical detection data show that the traditional Chinese medicinal preparation has clear advantages on aspects of improving signs and symptoms, improving liver funcotion, hepatitis B virus e antigen (HBeAg) negative conversion ratio, HBeAg/anti-HBe serum conversion ratio and hepatitis B virus desoxyribonucleic acid (HBV-DNA) negative conversion ratio.

The invention relates to the field of biotechnology, and particularly relates to a method for purifying type 71 enterovirus (EV71) from the cell culture harvest by a chromatographic process. The method comprises the following steps: clarifying the cell culture of the virus; performing ultrafiltration and concentration of the virus; purifying the virus through gel filtering chromatography and anion exchange chromatography. The continuous chromatography step has the advantages that the virus is purified and the replacement of chromatography buffer solution and adjustment of pH value and electrical conductivity are avoided; the purified virus liquid has the advantage of low residue of host protein and host desoxyribonucleic acid. The purified EV71 liquid prepared by the method can be used for preparing an inactivated vaccine against hand-foot-mouth diseases and can also be used for preparing reference antigen and antibody in a diagnostic reagent.