687 results about "RNA extraction" patented technology

The present invention provides methods and compositions for extracting RNA from cells. The cellular extract may be directly used in a variety of reactions, such as reverse transcription and PCR.

The invention discloses a nucleic acid extraction and purification method based on nanometer magnetic beads, comprising the following steps: mixing a biological sample and a lysis buffer to make nanometer magnetic beads in the lysis buffer and nucleic acid DNA / RNA which moves into the lysis buffer form a magnetic bead-nucleic acid compound; transferring the compound under the action of a magnetic field to a washing buffer to wash off impurities on the magnetic bead-nucleic acid compound; and transferring the washed magnetic bead- nucleic acid compound under the action of the magnetic field to an elution buffer so as to elute and recover nucleic acid. The nanometer magnetic beads used in the invention have advantages of uniform size, smooth surface, large surface area ratio, high adsorption capacity of nucleic acid, fast magnetic response speed and rapid separation, and can be stored together with the lysis buffer at room temperature for a long time. The extracted nucleic acid DNA / RNA has high purity, is complete and can be directly used for follow-up detection. The method provided by the invention has shorter nucleic acid extraction time than a general magnetic bead method by the use of a nucleic acid extraction reagent, is more suitable for automation and is adopted to realize high-flux nucleic acid DNA / RNA extraction.

Apparatus and method for collection of a target material from a liquid sample comprising target species that contain the target material. The apparatus comprises a fluid flow conduit in communication with a filter medium having a pore size adapted to retain target species thereon and pass, as filtrate, lysate containing the desired target material. A lysing agent conduit communicates with the fluid flow conduit and delivers lysing agent to the filter medium to lyse the target cells thereby releasing the desired intracellular target material. The lysing agent may be recirculated through the filter medium for a sufficient time to permit sufficient quantity of lysate to circulate through the system for lysate collection and subsequent assay.

The invention discloses a detection method for the expression of a GPR120 gene based on eGFP. The detection method comprises the following steps: inserting an eGFP fragment to the specific site TAA of the termination codon of the GPR120 gene by using CRISPR / Cas 9 technology so as to obtain a transgenic model mouse with eGFP-labeled GPR120 positive cells; and carrying out excitation by using a fluorescence analyzer and determining the fluorescence intensity of the positive cells of the mouse. The method carries out real-time monitoring on the expression of the GPR120 gene so as to control and reduce errors among groups, guarantee the reliability of results and compensate for and overcome the problems of variability caused by comparison of different groups of histocytes and incapability of monitoring the changes of gene expression at a living cell level in the prior art. According to the invention, the expression level of the GPR120 gene can be immediately determined after collection of cells, and operations and reactions like RNA extraction, inverse transcription and PCR are omitted, so detection of the expression of the GPR120 gene is simpler.

The invention relates to the field of molecular biology and discloses a method and a kit for extracting ribonucleic acid (RNA). The method for extracting the RNA comprises the following steps of: uniformly mixing a sample and lysis solution; centrifugally collecting supernate and uniformly mixing the supernate and absolute ethyl alcohol which is half of the volume of the supernate; combining the mixture with 0.45 mu m glass fiber cellulose acetate membrane; washing with deproteinization liquid and rinsing liquid; eluting adsorbed RNA; uniformly mixing lysis cells and an extraction aid; and centrifugally removing polysaccharide and polyphenol substances. The kit for extracting the RNA comprises the lysis solution, the deproteinization liquid, the rinsing liquid, the eluting liquid and the extraction aid. The method of the invention has the advantages of simpleness, rapidness, no use of toxic reagent, wide application range and capabilities of effectively removing the polysaccharide andpolyphenol substances and separating high-quality RNA out by adding the extraction aid. The kit of the invention has the advantages of no toxic reagent, wide application range, RNA extraction effectsof plant materials which are rich in polysaccharide and polyphenol superior to that of foreign kits, low cost and suitability for extensive laboratories and scientific researches.

The invention discloses a kit for extracting DNA / RNA of a virus by utilizing magnetic beads and a using method. The kit comprises the following eight components: an extracting solution I (cracking), an extracting solution II (binding), an extracting solution III (scrubbing), an extracting solution IV (scrubbing), an extracting solution V (eluting), magnetic bead suspension, a protease K working solution and a nucleic acid settling agent. The kit extraction method comprises the following steps: cracking, binding, scrubbing and eluting. The kit and the extraction method can be used for extracting the DNA / RNA of the viruses of different types of samples, the impurities such as proteins and lipids in the samples are effectively removed, the extracted nucleic acid is high in purity and complete in fragments, and the extraction process is safe and non-toxic.

The invention discloses an RT-LAMP (loop-mediated isothermal amplification) rapid detection kit for cucumber green mottle mosaic virus (CGMMV) and a detection method. The method comprises the steps of extracting plant viruses RNA, detecting an RT-LAMP reaction and electrophoresis detection or fluorochrome, and determining whether a plant carries the viruses. An RT-LAMP primer is designed according to a coat protein gene conserved region of CGMMV, and an RT-LAMP technology is adopted to establish a detection technology which can rapidly and sensitively detect the cucurbitaceous plant CGMMV such as cucumbers, watermelons and calabashes with high specificity and an application method thereof in diagnosing disease. The detection method provided by the invention can utilize the rapid extraction RNA and conventional RNA extraction as an RNA template for an RT-LAMP experiment. The method is utilized to rapidly identify a CGMMV plant sample, the targeted quarantine inspection protective measure is adopted, and the loss caused by the disease is reduced.

The present invention concerns the methods and compositions for preparing a tissue section or biological sample, particularly to preserve RNA in the section or sample, by not exposing or contacting the sample or section to a solution that is composed of mostly water. Tissue sections can be fixed, stained, and dehydrated for subsequent manipulation, including laser capture microdissection (LCM) for further analysis using methods and / or compositions of the invention.

The invention provides a kit for detecting 7 genetic markers of peripheral blood in early diagnosis of nasopharyngeal darcinoma. The kit comprises a peripheral blood total RNA extracting reagent, RT-PCR reaction liquid, time fluorescence quantitative RT-PCR reaction liquid and an SVM nasopharyngeal darcinoma diagnosis model, wherein the time fluorescence quantitative RT-PCR reaction liquid comprises primers of which the sequences are shown as SEQ IDNO:1-16; the primer sequences are SYBR Green fluorescence quantitative RT-PCR detection primers of the 7 genetic markers and reference gene GAPD; and the 7 genetic markers are HERC5, DLG7, PPARG, PLK1, KIF15, KLHL25 and MYST4. The invention also provides a using method for the kit. The kit has the advantages of high sensitivity, specificity and convenience, and can be used as aided diagnosis and early diagnosis technology for the nasopharyngeal darcinoma.

The present invention provides methods and compositions for extracting RNA from cells. The cellular extract may be directly used in a variety of reactions, such as reverse transcription and PCR.

The present invention provides RNA extraction reagents, methods and kits that are especially useful for extracting RNA. The reagents, methods and kits of the present invention are especially useful for extracting RNA, for example, cytoplasmic RNA, from difficult materials, from plants, especially, difficult plant tissues, such as those containing phenolics, tannins, polysaccharides (such as starch) and resins. Comparative high yields are obtainable according to the present invention when compared to conventional reagents and methods. The RNA preparations obtained in accordance with the present invention are also of high quality as demonstrated by superior A260 / 280 results and by gel electrophoresis.

The invention belongs to the field of molecular biology and particularly relates to a kit for extracting ribonucleic acid (RNA) by virtue of a paramagnetic particle method and an extraction method. The kit contains tissue digestion fluid, lysate, proteinase K, DNase I, DNase IBuffer, nucleic acid, extraction magnetic beads, washing liquid I, washing liquid II and eluant. The invention further discloses a method for extracting the tissue RNA by virtue of the kit. According to the kit and the method, the extraction yield and purity of RNA are increased, the integrity of RNA is improved, meanwhile, the automatic extraction is realized, and the simultaneous parallel testing of multiple samples is realized, so that the labor cost and the time cost are saved.

The invention relates to a preparation method of a silver nanometer membrane for detecting surface enhanced Raman spectra (SERS) of ribonucleic acid (RNA), and belongs to the technical field of medical science. In the method, the RNA in blood is extracted, and SERS spectra with strong signals and more spectrum peaks are acquired by taking the prepared silver nanometer membrane as an enhanced substrate to reflect the information of RNA molecules fully; and the enhanced substrate prepared by the method is contrasted to various enhanced substrates simultaneously to obtain the conclusion that the information acquired in the process of detecting the RNA molecules by using the enhanced substrate is the most and highest in strength, so high-quality SERS spectrograms can be acquired. Therefore, the high-quality spectrograms can be used for statistic analysis to obtain differences between patients and normal person so as to establish the foundation for the application of SERS detection to clinical diagnosis. The method has the advantages of short time for measurement and analysis and small using amount of samples, and is convenient and accurate in detection.

The invention discloses a method for extracting RNA from gymnosperm tissues. The method includes the following steps: 1) extracting buffer solution by RNA to treat plant materials; 2) protein removal: extracting by equal-volume chloroform / isoamylol for twice; 3) using LiCl to deposit RNA and SSTE to dissolve back RNA; 4) glycan removal: adding absolute ethyl alcohol and potassium acetate to the dissolved solution acquired in step 3) and depositing glycan in ice bath; 5) extracting chloroform / isoamylol again; 6) depositing RNA and DEPC H2O again to dissolve back RNA. In order to remove DNA, the following steps can be also conducted: a) using DNase without RNase pollution to remove DNA; b) using chloroform / isoamylol to extract and remove DNase; (c) repeating step 6). Wherein, RNA extract and buffer solution contains 100mmol / L Tris HCl(pH 8.0), 2g / 100mlCTAB, 2g / 100mlPVP, 2mol / LNaCl and 25mmol / L EDTA and is added with dithiothreitol with final concentration of 10mmol / L and 2 percent (volume percentage) of beta-mercaptoethanol. Therefore, the invention solves the interference of glycan and polyphenol and the like and acquires high-purity RNA.

The invention discloses an EML4-ALK fusion-gene non-invasive detection kit. A set of primers are provided and include EFTUD2-F1, EFTUD2-R1, EML4(13)-ALK-F2, EML4(20)-ALK-F3, EML4(6)-ALK-F4, EML4(6 / 13 / 20)-ALK-R, a detection probe ALK-EML4(6 / 13 / 20)-TAR, and a reference probe EFTUD2-REF. According to the EML4-ALK fusion-gene non-invasive detection kit, non-invasive plasma exosome is adopted to detectEML4-ALK fusion genes, the formula of an optimal RNA extraction agent is explored, the optimal RNA extraction agent serves as a template, and then digital PCR detection is conducted by the adoption of the found optimal primers and probes; the EML4-ALK fusion-gene non-invasive detection kit is high is detection sensibility, rapid, convenient and absolutely quantified, and guides drugs for the EML4-ALK fusion genes.

The invention relates to the field of nanometer materials, and discloses a silicon dioxide nanofiber membrane, and a preparation method and application thereof. The method comprises the following steps: (1) preparing spinning solution containing nanosilicon dioxide, a surface active agent, a high-molecular polymer and solvent, and (2) processing the spinning solution prepared in step (1) by utilizing electrospinning technique, and drying to obtain the silicon dioxide nanofiber membrane, wherein the average grain diameter of the nanosilicon dioxide in step (1) ranges from 20nm to 120nm. The fiber diameters of the silicon dioxide nanofiber membrane which is prepared by the method provided by the invention range from 80nm to 600nm; the thickness is small; therefore, the bore diameter and specific surface area of the silicon dioxide nanofiber membrane are relatively large; the surface energy and surface activity are high; the silicon dioxide nanofiber membrane provided by the invention can be applied into an RNA (ribose nucleic acid) extraction kit adsorbing column, and has the advantages that the extraction scope and category are wide, the extraction is rapid, simple and convenient, the purification rate is high, and the ratio of performance to price is high.

The invention provides a detection method of Chikungunya virus, which comprises the procedures as follows: pretreatment of test sample; RNA extraction of test sample to be detected; design and synthesis of primer and probe; determination of optimum fluorescence quantitative PCR reaction system; program amplification and adjudging results according to Ct value. The optimum fluorescence quantitative PCR reaction system is determined as follows: the CHIK-FP terminal concentration of positive primer is 500nM, the CHIK-RP terminal concentration of reverse primer is 1000nM, and the CHIK-Probe terminal concentration of probe is 250nM. The amplification program is 45 times of circulation of 10 minutes at 50 DEG C, 10 minutes at 95 DEG C, 10 seconds at 95 DEG C and 30 seconds at 62 DEG C. The invention establishes a detection method for Chikungunya virus, which can be applied on monitoring of the virus, and can further strengthen the monitoring work for preventing from spreading the disease into our country and has important social efficiency and economic efficiency.

The invention relates to a BCR / ABL fusion gene (P210bcr / abl) mRNA fluorescence quantitative PCR measurement detection reagent box which comprises lymphocyte segregating liquid, RNA extracting liquid A, RNA extracting liquid B, Oligo (dT) 12-18, RT reaction fluid, reverse transcriptase, quantitative PCR reaction fluid, standard sample, comparison sample, and DEPC water; wherein, a marked primer and a non-marked primer are contained in the quantitative PCR reaction fluid. The reagent box can accurately detect the BCR / ABL fusion gene (P210) mRNA level in the specimen to be detected by extracting the total RNA of medulla ossium or peripheral blood, obtaining cDNA through reverse transcriptase and being combined with a real-time fluorescence quantitative PCR measurement detection technology. The reagent box adopts the latest self-quenched probe technology, thereby having the advantages that the repetitiveness is good, the sensitivity is high, the cost is low, and the invention can be applied to the dynamic monitoring of chronic myelocytic leukemia diagnosis, curative effect observation, prognosis and micro residual leukemia (MRD).

The invention provides a method for extracting DNA and RNA of a plant synchronously, which utilizes one plant sample; and after the plant sample is treated and decontaminated with the same steps, the DNA and RNA thereof are settled and purified respectively, thereby achieving the purpose of extracting synchronously. The method can be applied to the synchronous extraction of DNA and RNA of a yam sample and has the advantages of being able to synchronously conduct nucleic acid extraction, time-saving, economical and fast; besides, the ultracentrifugation, KAC and the like adopted by the method are very effective in eliminating the impurities contained in the yam such as amylose and the like; the DNA / RNA extraction quality of the method is high; the RNA OD260 / 280 ratio is between (1.8 to 2.0) and the DNA is between 1.7 to 1.8; and the electrophoretic band is complete and clear. The method is also applicable to the nucleic acid extraction of other plant samples.

The detection process of SARS virus of the gene detection reagent kit with rationing standard specific SARS virus nucleic acid as reference includes RNA extraction, PCR amplification and fluorescent detection. The venous blood sample, gargled liquid or respiratory tract secretion of the patient is used as the analysis sample directly, RNA of the sample is extracted as the nucleic acid template, and the template is used in fluorescent PCR amplification. The used fluorescent amplification detecting reagents includes one-step process RT-PCR buffering liquid, deoxynucleoside triphosphate (dNTP) mixture, specific amplification primer and specific probe. The method of the present invention is fast and convenient in detecting SARS virus.

The invention relates to a porcine reproductive and respiratory syndrome virus RT-LAMP detection kit and a detection method thereof. Primers required by PRSSV RT-LAMP reaction system are designed according to the sequence of porcine reproductive and respiratory syndrome virus (PRSSV) published by GenBank; PRSSV virus RNA is extracted with the virus RNA extraction reagent (LBBII-RNA) designed and prepared by the inventor, the PRSSV RT-LAMP reaction system established in the invention is utilized for detection, and color developing agent is added after the reaction to judge the result; the result shows that the PRSSV virus RNA obtains efficient specific amplification after the reaction is conducted for 45 minutes at the temperature of 63 DEG C; and then, quick detection of porcine reproductive and respiratory syndrome virus American classical strain and NSP2 variant strain (highly pathogenic porcine reproductive and respiratory syndrome virus strain) is conducted by SpuI enzyme cutting. Compared with the prior art, the invention has quick detection, high sensitivity, low reaction cost, convenient and fast operation, which is capable of differentiating American classical strain and NSP2 variant strain (highly pathogenic porcine reproductive and respiratory syndrome virus strain) and meets the requirement of multi-level detection.

The invention discloses a universal primer and probe combination for detecting foot and mouth disease viruses through RPA (recombinase polymerase amplification)-lateral flow assay technology. The forward primer sequence is shown as SEQ ID No.1, the reverse primer sequence is shown as SEQ ID No.2, and the probe sequence is shown as SEQ ID No.3. The invention also discloses a kit for detecting foot and mouth disease viruses. Detection with the primers and probes involved in the invention only needs virus RNA extraction on clinical samples and isothermal amplification after one-step process reverse transcription into cDNA, thermal cycling reaction is not needed, amplification in a PCR instrument is unnecessary, and the result can be clearly displayed on a lateral flow chromatography test strip. The primer and probe combination and the kit provided by the invention have the advantages of high sensitivity, strong specificity, simple reaction procedure, short detection time and the like, and are suitable for rapid, accurate and simple detection of foot and mouth disease viruses on a cattle farm.

The invention discloses a digital PCR-based novel coronavirus nucleic acid quantitative detection kit and an application. The reaction total system of the digital PCR-based novel coronavirus nucleic acid quantitative detection kit has 20 ul, comprising 10ul of 2x One-Step RT-ddPCR Supermix, 0.8ul of 25mM manganese acetate solution, 5ul of to-be-detected sample RNA, 1ul of ORFlab gene primer probeworking solution, 1ul of N gene primer probe working solution, 1ul of RPP30 gene primer probe working solution and 1.2ul of Nuclease-Free Water, wherein 5ul of negative control RNA extraction solutionand 5ul of positive control RNA extraction solution are adopted to replace the to-be-detected sample RNA in a negative control reaction system and a positive control reaction system respectively. Based on the innovative RNA one-step reverse transcription microdroplet type digital PCR technology, nucleic acid absolute quantification is carried out specific to highly conservative ORFlab gene and Ngene in the novel coronavirus (2019-nCoV) genome, so that the detection accuracy is improved, and the kit can be used for clinical assisted diagnosis and viral load analysis of novel coronavirus (2019-nCoV) infection, and has a wide clinical application value.

The invention discloses a detection kit and detection method for H1, H3 and H9 type avian influenza viruses and belongs to the technical field of biology. The kit comprises an RNA extraction kit, an RNA reverse transcription kit and an RPA detection kit. The RNA extraction kit is used for extracting RNA of a to-be-detected sample to be used as a detection template. The RNA reverse transcription kit is used for conducting reverse transcription on the RNA in the detection template to obtain cDNA. The RPA detection kit comprises primers and fluorescent dye. The primers are used for amplifying the cDNA to obtain an amplified DNA sequence. The primers comprise the forward primer and the reverse primer. The fluorescent dye is used for conducting fluorescence detection on the amplified DNA sequence. According to the detection kit and the detection method, the avian influenza viruses can be rapidly, easily and conveniently detected.

The invention discloses a kit for simultaneously detecting nucleic acids of seven respiratory pathogens and application of the kit. The kit is based on a double amplification technology (RNA isothermal amplification and multi-biotin signal amplification), and can detect H1N1 / H3N2 type influenza A viruses, influenza B viruses, respiratory syncytial viruses, 1 / 2 / 3 type human parainfluenza viruses, B / E type adenoviruses, mycoplasma pneumoniae and chlamydia pneumoniae. The kit of the invention does not need RNA extraction, and is not prone to pollution, high in sensitivity and good in specificityin the process of detection, and can be widely applied to detection of the nucleic acids of the above seven respiratory pathogens.

The invention discloses a method for extracting RNA of sweet potato root tuber and application thereof. Beta-mercaptoethanol and polyvinylpyrrolidone are used to prevent polyphenols from interfering the separation of RNA; and in addition, the NaCl with high concentration, absolute ethyl alcohol and potassium acetate are used, so the polysaccharose can be rapidly precipitated, and the influence of the polysaccharose on the extraction of RNA can be prevented. When preventing the interference effect of the polyphenols and the polysaccharose on the separation of RNA, RANA can still be remained in the solution, so the RNA with good quality, high purity and good completeness can be obtained, and the attainment rate and the yield of the RNA are large. The method has high practical application value, and can satisfy the research of molecular biology such as reverse transcription, Northern hybridization, cDNA library construction, transcriptome analysis and the like.

The invention discloses a primer and a probe combination for detecting bovine viral diarrhea virus by a recombinase polymerase application (RPA) technology. The forward primer sequence is as shown in SEQ ID No.1; the reverse primer sequence is as shown in SEQ ID No.2; and the probe sequence is as shown in SEQ ID No.3. The invention also discloses a kit for bovine viral diarrhea virus detection. When the primer and the probe are used for detecting, a clinical sample only needs the treatment steps of virus RNA extraction, reverse transcription by a one-step method to form cDNA, and isothermal amplification, thermal cycle reaction is not needed, amplification does not need to be performed in a polymerase chain reaction (PCR) instrument, and the result can be displayed clearly on a lateral flow assay test strip, so that the primer and the probe have the advantages of high sensitivity, high specificity, simple reaction procedure, short detection time and the like, and are applicable to quick, accurate, simple and convenient diagnosis and treatment on the diseases in cattle farms.

The invention discloses a fluorescent reverse transcription-polymerase chain reaction (RT-PCR) kit for quantitatively detecting a leukemia translocation ETS leukemia acute myelocytic leukemia (TEL-AML)1 fusion gene and belongs to the field of in-vitro nucleic acid diagnosis. The kit comprises a quantitative reference substance, a negative reference substance, a positive reference substance, RT-PCR enzyme, PCREnhancer, diethypyrocarbonate (DEPC) water, fluorescent PCR reaction liquid I, fluorescent PCR reaction liquid II and ribose nucleic acid (RNA) extraction liquid. The kit comprises an RT-PCR system in which a fluorescent PCR technology is taken as basis, and forward and reverse primers and a fluorescent probe aiming at AMLI and TEL-AML1, can detect the RNA of AMLI and TEL-AML1 simultaneously under the same PCR condition, detect whether TEL-AML1 gene fusion occurs in a clinical sample conveniently and quickly, and provides important basis for leukemia diagnosis, typing, clinical treatment and prognosis diagnosis, and a new train of thought for clinical treatment.

The invention provides a method for extracting, purifying and detecting RV RNA (Rubella Virus Ribose Nucleic Acid), and a corresponding kit for detecting the RV RNA. The kit comprises a RNA extracting solution containing magnetic beads, and a PCR (Polymerase Chain Reaction) reaction liquid containing an upstream primer, a downstream primer and a probe, wherein the upstream primer and the downstream primer are used for amplifying target polynucleotides; the probe is used for detecting the target polynucleotides. The kit can be used for detecting the RV RNA and cannot be used for detecting non-RV pathogens, thereby illustrating that the kit has the good specificity. In addition, the RNA is extracted by selecting a method of the magnetic beads which are good in adsorption effect and easy to purify, so that the RNA with a high purity and a high yield can be obtained. Thus, the detection sensitivity, the detection accuracy and the detection stability of the kit are greatly improved, wherein the lower limit of detection of the RNA, namely, the sensitivity of the kit can reach 400copies / ml; the detection range of the kit (the quantitative linear range of the kit) can reach 4.00E+02copies / ml to 4.00E+08copies / ml.