63results about How to "Optimize the reaction system" patented technology

The invention discloses a construction method of a small-fragment DNA (deoxyribonucleic acid) library based on an Illumina Hiseq 2500 sequencing platform, which comprises the following steps: blood free DNA extraction, fragment size screening, terminal repair, 3' terminal linker addition, PCR (polymerase chain reaction) amplification, magnetic bead purification and the like. Compared with the library construction process based on a sequencing platform in the past, the method disclosed by the invention simplifies the experiment process, shortens the library construction time, optimizes the reaction system, greatly reduces the reagent consumption, lowers the library construction cost, lowers the library loss, and is suitable for constructing an artificially-fragmented small DNA library.

The invention discloses a detecting method for leptospira, which is based on the quantitative detection technique of nucleic acid isothermal expansion, which is characterized in that a specific primer is used, and the specific area of target gene is expanded utilizing the LAMP technological platform; under the assistance of a series of quality control and internal control detection system, the leptospira is detected in molecule level, which can realize the qualitative and quantitative detection to micro sample. Compared with the prior art, the detecting method for leptospira has the advantages of decreased non-specific expansion because of isothermal expansion, good specificity, fast detection speed, high sensitivity, no need for special equipment, low cost, easy operation, easy popularization and wide application prospect, and is suitable for experimental study, clinical detection and environment detection.

The invention relates to a diarrhea pathogen multi-gene detection system as well as a kit and application thereof. The detection system comprises 22 pairs of primers, wherein the 22 pairs of primers comprise 19 pairs of diarrhea pathogens, 2 pairs of human genome internal references and 1 pair of system quality control internal reference primers. The diarrhea pathogens are used for treating campylobacter jejuni, shigella, clostridium difficile, salmonella enteritidis, salmonella typhimurium, enterotoxin of escherichia coli, escherichia coli O157, vibrio, yersinia enterocolitica, human astrovirus, norovirus II, human enteric adenovirus, rotavirus and the like. According to the diarrhea pathogen multi-gene detection system and the kit thereof, steps including conventional culture and the like do not need to be adopted and synchronous detection and analysis of a plurality of types of the diarrhea pathogens can be directly carried out on an excrement sample in the same reaction system; the disadvantages of a conventional detection method that the flux is low, the consumed time is long, the detection rate is low and the like are made up; comprehensive, precision and low-cost pathogen diagnosis is provided for clinic at the first time and important reference is provided for individualized drug administration and precision medical treatment.

The embodiment of the invention discloses a nucleic acid fluorescence PCR detection kit for a universal enterovirus, a coxsackievirus A16 and an enterovirus 71. The nucleic acid fluorescence PCR detection kit comprises an RNA extracting solution, an RNA eluant, internal standards, a PCR reaction solution, an EV / CA16 / EV71 enzyme mixed solution, EV / CA16 / EV71 positive reference substances and EV / CA16 / EV71 negative reference substances. According to the kit, the universal enterovirus, the coxsackievirus A16 and the enterovirus 71 can be simultaneously detected in a same sample, but other pathogen RNA cannot be detected, the detection sensitivity can reach 400 copie / ml, the detection range is 4.0 E+02-4.0E+08 copies / ml, and a reliable experimental evidence is supplied to early diagnosis on infection of the universal enterovirus, the coxsackievirus A16 and the enterovirus 71.

This disclosure concerns the systems, methods, and kits for the in vitro synthesis of biological macromolecules in a reaction utilizing cell lysates containing plastids, mitochondria and / or chloroplasts, wherein creatine phosphate and creatine kinase are not added to the reaction to provide artificial energy regeneration.

The invention provides a loop-mediated isothermal amplification test method of an influenza A3 virus. The method comprises the following steps: a specificity primer sequence is designed according to an influenza A3 virus hemagglutinin gene, a sample RNA to be tested is extracted as the template to have LAMP reaction; the LAMP reaction result is analyzed; if the LAMP amplification result is positive, the sample to be tested contains the influenza A3 virus. The LAMP influenza A3 virus test method is characterized by convenience, economy, fastness, sensitivity and specificity with flexible and convenient result judgment, is applicable in clinical or laboratory diagnosis, and has broad application prospect.

The invention relates to a multiple-gene detecting kit related to antitumor drugs. The multiple-gene detecting kit comprises a mixture of RT (reverse transcription) primers and a mixture of PCR (polymerase chain reaction) amplification primers, wherein each RT primer and each PCR amplification primer based on genetic groups, reference genes and a reaction internal label are respectively contained in each of the mixture of the RT primers and the mixture of the PCR amplification primers; the multiple-gene detecting kit is characterized in that the genetic groups comprise PTEN, EGFR, DPYD, HER2, RRM1, ERCC1, TUBB3, TOP1, TYMS and TOP2A; the reference genes comprise ACTB, GAPDH and B2M; the reaction internal label is KAN-rRNA (ribosomal RNA). The multiple-gene detecting kit related to antitumor drugs disclosed by the invention can be used for systemically detecting the expression level of a plurality of genes closely related to the antitumor drugs by one step, is simple and convenient to use, good in accuracy and high in detecting efficiency, and can be used for guiding chemotherapy drugs and selecting chemotherapy regimens very well.

The invention provides a homocysteine detecting kit and a using method thereof, relates to the field of biochemical detection, and aims to provide a homocysteine detecting kit which is high in stability, high in accuracy and high in precision and a using method of the homocysteine detecting kit. The homocysteine detecting kit comprises a reagent 1 and a reagent 2, wherein the reagent 1 comprises a capso buffer solution, lactic dehydrogenase (LDH), L-serine, reduced coenzyme Thio-NADH, tris((2-carboxyethyl)phosphine, mercaptoethanol, an alkylphenol polyoxyethylene series, EDTA, polyethylene glycol 300, ascorbic acid oxidase, a composite stabilizer, and a proclin series preservative; and the reagent 2 comprises a capso buffer solution, cystathionine-beta-synthase, methionine synthase, cystathionine-beta-lyase, methionine-gamma lyase, a surfactant and sodium dodecylbenzene sulfonate. By the various added mixed enzymes and methionine-gamma lyases, the accuracy of the kit on detection of homocysteine can be improved.

The invention discloses a method for preparing dichlorofluoroethane by catalytic fluorination with vinylidene chloride. The method comprises the following steps: (1) preparing vinylidene chloride, namely preparing a carbon nitride catalyst, additionally chlorinating vinyl chloride and hydrogen chloride to generate 1,1-dichloroethane, and re-chlorinating the 1,1-dichloroethane to generate 1,1,1-trichloroethane; adding the carbon-nitride catalyst into the 1,1,1-trichloroethane, reacting at 550 DEG C to obtain vinylidene chloride, cooling, sealing and storing; (2) preparing anhydrous hydrogen fluoride; and (3) synthesizing dichlorofluoroethane, namely adding vinylidene chloride and anhydrous hydrogen fluoride into an ultrasonic oscillator for mixing, performing ultrasonic oscillation for 30 minutes to obtain a reacting liquid, putting the reacting liquid into a reaction kettle, and reacting under bluelight catalysis to generate the dichlorofluoroethane. The method can be used for effectively inhibiting production of byproduct dichlorofluoroethane, and has high vinylidene chloride conversion rate and high synthesizing speed, and the production efficiency can be improved.

The invention discloses a method for identifying the germplasm resource of radix codonopsis by use of ISSR (inter-simple sequence repeat) fingerprint. The method comprises the following steps: (1) collecting and numbering radix codonopsis from different production places; extracting the DNA of the radix codonopsis from different production places and performing PCR amplification; detecting amplification products by agarose gel electrophoresis; according to the result of agarose gel electrophoresis and by adopting the number of radix codonopsis as a horizontal coordinate and the length of DNA fragment as a longitudinal coordinate, recording the detection results of the DNA amplification stripes of the radix codonopsis from different production places at the intersections between the horizontal coordinate and the longitudinal coordinate respectively; marking the detection result with DNA amplification stripe, and not marking the detection result without DNA amplification stripe, so as toobtain the ISSR fingerprints of the radix codonopsis from different production places; (2) comparing the ISSR fingerprint of the to-be-detected radix codonopsis with the ISSR fingerprints of the radix codonopsis from different production places, and judging the production place of the to-be-detected radix codonopsis according to the comparison result. The method disclosed by the invention can beused for conveniently and quickly identifying the radix codonopsis medicinal materials with little difference in appearance and easily confused, and quick and accurate identification can be performedin the seedling stage of the radix codonopsis; therefore, the method is of great significance on guaranteeing the accuracy and reliability of the base source of a medicinal material.

The invention relates to the field of a serum fibronectin detection technology, in particular to a fibronectin detection reagent. A reagent R 1 contains buffer solution, gamma-Fe2O3 / polythiophene (gamma-Fe2O3 / PTP) composite nanoparticles, fluorocarbon surfactant FC-4430 and preservative; a reagent R 2 contains buffer solution, goat anti human IgG fibronectin antibody, gelatin, xanthan glue, sodium chloride, mycose, PEG-6000, fluorocarbon surfactant FC-4430 and preservative. According to the fibronectin detection reagent, the reacting system is optimized, the stability of the reagent can be improved significantly through the adoption of HEPES (4-hydroxyethylpiperazine ethane sulfonic acid) buffer solution and the addition of a variety of stabilizers, such as sodium chloride, mycose, PEG-6000, gelatin and xanthan glue; the addition of gamma-Fe2O3 / polythiophene (gamma-Fe2O3 / PTP) composite nanoparticles in the reagent can adsorb the antigen-antibody complex through electrostatic adsorption during the detection, thus the sensitivity and accuracy degree of the reagent can be enhanced significantly. Moreover, the optimized novel fluorocarbon surfactant FC-4430 can promote and maintain the stability of the antibody, prevent the system from being muddied, and further enhance the stability of the reagent.

The invention provides a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) reagent for detecting 2019-nCov and application of the CRISPR reagent in preparation of a kit for detectingthe 2019-nCov. The CRISPR reagent comprises an RPA (Recombinase Polymerase Amplification) primer pair and corresponding crRNA. Based on the CRISPR molecular diagnosis technology, through coupling of RPA-CRISPR, two-stage amplification of 'sequence amplification' (RPA completion) and 'enzymatic cascade' (Cas enzyme completion) can be realized, so that 2019-nCov is detected with high sensitivity andhigh specificity under the condition of a simple instrument (metal bath).

The invention relates to a diarrhea pathogenic bacteria multi-gene detection system and its kit and application. The detection system has 15 pairs of primers, wherein 13 pairs of diarrhea pathogens primers, one pair of human genome beta-actin primers and one pair of system quality controlled beta-actin primers are included. Diarrhea pathogens refer to campylobacter jejuni, shigella, clostridium difficile, salmonella enteritidis, salmonella typhimurium, enterotoxigenic escherichia coli, escherichia coli O157, vibriones, yersinia enterocolitica and the like. The diarrhea pathogenic bacteria multi-gene detection system and its kit do not require routine culture and other steps, synchronous detection and analysis of the various diarrhea pathogens can be directly conducted on a stool sample in the same reaction system, the shortcomings that a routine detection method is low in flux and low in detection rate and takes much time are made up for, a comprehensive, accurate and low-cost pathogen diagnosis is provided for clinical use for the first time, and an important reference is provided for individualized medication and accurate medical treatment.

The invention discloses a composition, a kit and a method for rapidly detecting mycoplasma ovipneumoniae. The accurate detection of mycoplasma ovipneumoniae is completed by designing specific primers and probes of ketolase gene of mycoplasma ovipneumoniae. The best reaction time and the best reaction temperature in the detection process, the specificity, sensitivity, repeatability and stability of the detection are all explored, the best reaction condition is found, and a rapid detection method of mycoplasma ovipneumoniae with good specificity, high sensitivity and stable repetition is established; and the method is simple to operate, convenient and time-saving, does not need large-scale experimental instruments and equipment, is very suitable for personnel without any experimental basis to operate, and realizes on-site detection.

A method for producing a melamine salt of bis(pentaerythritol phosphate) phosphoric acid is proposed. The method includes the steps of reacting and mixing pentaerythritol and diphosphorous pentaoxide in an extruder to carry out esterification to produce bis(pentaerythritol phosphate) phosphoric acid; and mixing bis(pentaerythritol phosphate) phosphoric acid with melamine or a derivative thereof to carry out quaternization in the presence of a solvent, followed by removing water to obtain a melamine salt of bis(pentaerythritol phosphate) phosphoric acid. Diphosphorous pentaoxide is used in the method as a reactant, so that no hydrogen chloride is produced and it is not necessary to recollect trichlorophosphoric acid. Moreover, in the method of the present invention, such that esterification time can be shortened and a high pressure in a closed reaction system that causes explosion can be avoided.

The invention discloses a primer composition and detection method for detecting novel coronaviruses, and particularly discloses a method for detecting novel coronaviruses by a loop-mediated isothermalamplification technique. An ORF1ab gene and an N gene in a complete genome of the novel coronaviruses are used as target genes, a specific primer is designed, and a reaction system is optimized, so that the target genes are amplified. The primer composition has the characteristics of being simple to operate, high in efficiency and low in cost. The primer composition has the characteristics of being high in specificity and convenient and swift when being used for detecting the novel coronaviruses, the detection efficiency can be improved, the detection requirements of a basal laboratory can bemet, and a new method is provided for detection of the novel coronaviruses.

The invention pertains to reaction systems in which at least one capillary unit is applied. The capillary unit comprises: —a unit inlet for receiving a fluid flow, —a unit outlet for releasing said fluid flow, —a capillary, which capillary is arranged between the unit inlet and the unit outlet such that said fluid flow passes through the capillary, —a heater and / or a cooler for adjusting the temperature of the capillary and therewith influencing the flow rate of the fluid flow passing through said capillary, —a housing for accommodating at least the capillary and heater and / or cooler of said capillary unit, which housing provides thermal insulation of the capillary, —a flow sensor for measuring the flow rate of the fluid flow through the capillary unit, which flow sensor can be arranged either inside or outside the housing, A capillary unit is combined with or further has a flow adjustment unit for adjusting the flow rates of the secondary fluid flows, which flow adjustment unit comprises a temperature control device for individually controlling the heater and / or cooler of each capillary unit in response to the flow rate that is measured by the flow sensor of that capillary unit.

The invention relates to a multi-gene detection kit for the selection of chemotherapy regimens. The multi-gene detection kit comprises a mixture of RT (reverse transcriptase primer) primers and a mixture of PCR (Polymerase Chain Reaction) amplification primers, wherein the mixture of RT primers and the mixture of PCR amplification primers comprise each RT primer and each PCR amplification primer based on genetic groups, reference genes and an internal standard substance of the reaction respectively, also comprise random primers, the genetic groups comprise RRM1, TOP2A, ERCC1, TYMS, TUBB3, TOP1, PTEN, HER2, DPYD and EGFR; the reference genes comprise ACTB, GAPDH and B2M; the internal standard substance of the reaction is KAN-rRNA. The multi-gene detection kit for the selection of chemotherapy regimens, which is disclosed by the invention, can be used for simultaneously detecting the expression levels of multiple genes closely related to anti-tumor medicines systematically, and is easy to use, good in accuracy, high in detection efficiency and can be well used to guide chemotherapy medication and select chemotherapy regimens.

The invention discloses primers, probes and kit for synchronously detecting human herpesvirus-6 and human herpesvirus-7. According to the primers, the probes and the kit, through selecting a novel target gene and redesigning the primers and the probes, the reaction system is optimized, the false positive incidence of clinical synchronous detection on human herpesviruses is substantially lowered, the sensitivity is high, the specificity is good, and the time for detection is shorter, so that the clinical application and popularization of the technology can be achieved.

The invention relates to a multiple nucleic acid detection system, which comprises an amplification primer group and a detection probe group aiming at a target nucleic acid sequence, the detection system comprises a Target probe modified by LNA and a Beacon probe for achieving a fluorescence quenching effect through 1-8 continuous G basic groups, and meanwhile, based on the proposal of the multiple nucleic acid detection system, the inventor combines a touchdown PCR (Polymerase Chain Reaction) program to detect the target nucleic acid sequence. The invention further provides a multiple nucleic acid detection method which can further reduce non-specific amplification in PCR reaction and improve detection sensitivity. Therefore, the limitation of traditional real-time fluorescent quantitative PCR typing is overcome, single-tube multi-typing is realized through special signal and melting curve analysis, and accurate qualitative detection of each target gene in at most 20 to-be-detected target nucleic acid sequences in a sample can be realized with a simpler reaction system and lower detection cost.

The invention discloses a primer, a probe and a kit for synchronously detecting human herpes virus types 6, 7 and 8. By selecting a new target gene, redesigning a primer and a kit, and optimizing a reaction system, the false positive rate of clinical synchronous detection on human herpes viruses is greatly reduced, the primer, the probe and the kit are good in sensitivity, good in specificity and short in detection time, and the technique can be applied to clinical application and popularization.

The invention discloses a primer for detecting FHV-1, a nucleic acid capture gold-labeled test strip, a reagent kit and an application. A primer pair for detecting FHV-1 comprises a first primer and asecond primer with sequences shown as SEQ ID NO. 7 and SEQ ID NO. 8 respectively. The gold-labeled test strip comprises a lining plate, wherein a sample pad, a gold-labeled combining pad combined with an FITC gold-labeled antibody, a coating membrane and a water absorption pad are sequentially arranged on the upper end surface of the lining plate in a set direction; an invisible detection area combined with a digoxin antibody and an invisible control area combined with a sheep anti-rat secondary antibody are arranged on the coating membrane. The reagent kit comprises the primer pair and the nucleic acid capture gold-labeled test strip. A detection method of FHV-1 is high in specificity and sensitivity, accurate and quick, and has wide application prospects.

The invention provides a rapid detection kit for snake source components and application of the rapid detection kit, and relates to the technical field of biological species identification. Accordingto the rapid detection kit, a snake source general internal standard gene JUN (Gen bank No. EF144048.1) is screened out firstly, the nucleotide sequence of the snake source general internal standard gene JUN is shown as SEQ ID NO.1, the snake source general internal standard gene JUN is located on a chromosome, the copy number of the snake source general internal standard gene JUN in snake speciesis constant, allele variation does not exist, and the snake source general internal standard gene JUN can serve as a target gene for identifying a snake source. A constant-temperature amplification primer is designed with the snake source general internal standard gene JUN as a target sequence, and a constant-temperature amplification reaction product is used for colloidal gold nucleic acid teststrip detection. The rapid detection kit is formed by combining constant-temperature amplification reaction with colloidal gold nucleic acid test strip detection, the snake source components can be rapidly and sensitively detected, and the detection sensitivity can reach 0.8% (w / w). The rapid detection kit is simple in using method, low in cost, good in specificity and quite suitable for on-site real-time detection, and a reaction result is easy to observe.

The invention discloses a nucleotide sequence for detecting bovine enterovirus and foot-and-mouth disease virus, and the nucleotide sequence for detecting the bovine enterovirus and the foot-and-mouth disease virus is as shown in SEQ (sequence) ID (identity) NO: 1 to SEQ ID NO: 4 in a sequence table, wherein the SEQ ID NO: 1 and the SEQ ID NO: 2 are a sense primer and an antisense primer for detecting the bovine enterovirus respectively; and the SEQ ID NO: 3 and the SEQ ID NO: 4 are the sense primer and the antisense primer for detecting the foot-and-mouth disease virus respectively. The invention further discloses a kit containing the primers for detecting the bovine enterovirus and the foot-and-mouth disease virus. The kit has the characteristics of quickness, sensitiveness, specificity, stability and good repeatability.

The invention discloses peptide nucleic acid of a UGT1A1 gene polymorphism detection primer, and the nucleic acid comprises a forward primer UGT1A1-F1, a reverse primer UGT1A1-R1, a forward primer UGT1A1-F2, a reverse primer UGT1A1-R2 and peptide nucleic acid, wherein the nucleotide sequence of the forward primer UGT1A1-F1 is as shown in the SEQ ID NO.1; the nucleotide sequence of the reverse primer UGT1A1-R1 is as shown in the SEQ ID NO.2; the nucleotide sequence of the forward primer UGT1A1-F2 is as shown in the SEQ ID NO.3; the nucleotide sequence of the reverse primer UGT1A1-R2 is as shown in the SEQ ID NO.4; the nucleotide sequence of the peptide nucleic acid is as shown in the SEQ ID NO.5; and the 5' ends of the reverse primer UGT1A1-R1 and the reverse primer UGT1A1-R2 are provided with fluorescence marks. The UGT1A1 gene polymorphism detection kit disclosed by the invention has high sensitivity, low template amount, high speed, accuracy and low cost.

The invention discloses a preparation method and an application of uracil DNA glycosidase. The uracil DNA glycosidase prepared by using the method can be used for hydrolyze glucosidic bonds between uracil basic groups and deoxyriboses in DNA at the temperature of 20-37 DEG C; at the same time the glycosidase has a good heat sensitivity, when the temperature is higher than 60 DEG C, the glycosidase swiftly loses activeness, when the temperature reaches 60 DEG C for 5 minutes, the enzymatic activity loses by 99%; the prepared heat sensitive uracil DNA glycosidase can be used for preventing the sample DNA contamination in all types of nucleic acid detection reaction. The preparation method and application of uracil DNA glycosidase have the advantages being capable of both completely removing the contaminated DNA, and exerting no inhibition to the nucleic acid amplified reaction. The glycosidase can be used as a cleaning agent to the sample contamination in all types of nucleic acid amplified reaction (such as PCR), can remove the DNA contamination caused by the products of the previous amplified reaction, and increase the accuracy of nucleic acid test.

The invention provides a plasminogen immunoturbidimetry detection kit. Trehalose is added into a reagent R1, so that the reaction system is more viscous, and relatively neutral, and the problem that the antibody is not stable in a dilute solution is solved to a certain extent, thereby playing a certain role in protecting the antibody; meanwhile, goat anti-human Pg antibody coated latex particles,instead of a goat anti-human Pg antibody, are added into a reagent R2, so that the antibody is protected fundamentally. Therefore, by virtue of the joint effect of the trehalose and the latex particles, the stability of the kit is effectively enhanced, influence to the accuracy of the reagent is avoided, and further popularization of the reagent in the market is promoted.

The invention provides a set of primers for detection of a porcine Japanese encephalitis virus. The primers include: an upstream internal primer, a downstream external primer, an upstream external primer, a downstream external primer, an upstream loop primer and a downstream loop primer. The sequences of the primers are shown as SEQ ID No.1-6 in a sequence table. The invention also provides a visual kit that contains the set of primers and is used for detecting the porcine Japanese encephalitis virus and a method for detecting whether a non-living sample contains the porcine Japanese encephalitis virus by the kit. The primers, the kit and the detection method provided in the invention optimizes the reaction system, and make the result determination realize visualization, thus being of great significance in detection, diagnosis, prevention and treatment of porcine Japanese encephalitis.

The invention discloses a method and kit for directly amplifying a DNA (deoxyribonucleic acid) bar code of a trace sample without extraction. The method comprises the following steps: adding a lysis solution and a buffer solution in a volume ratio of 1:(0-9) to obtain a trace sample DNA solution, and carrying out (polymerase chain reaction) amplification by using a forward primer LCO1490 and reverse primer HCO2198 to obtain the DNA bar code sequence of the trace sample. By optimizing the formula and the ratio of the lysis solution and the buffer solution and optimizing the reaction system and reaction conditions, the invention can avoid mixed extraction of template DNAs on a plurality of mini animal individuals, thereby ensuring the singleness of the DNA template source. The method disclosed by the invention has the advantages of high favorable, high amplification efficiency, high reliability and wide adaptability.

The invention discloses an rs12979860 locus genotyping dual-color fluorescent PCR rapid detection kit. According to the kit, a primer and a TaqMan-MGB probe are redesigned, a reaction system is optimized, bi-component hot-start DNA polymerase forms an enzyme activity automatic regulation system, ROX Reference Dye can eliminate a signal background and correct fluorescence signal errors between holes, therefore, the kit achieves accuracy, high amplification efficiency, high sensitivity, good specificity, good repeatability, easy and convenient operation and shorter detection time, and the technology can be applied and popularized clinically.