44results about How to "Quantitative results are accurate and reliable" patented technology

The invention relates to a real-time fluorescence quantitative PCR (polymerase chain reaction) kit for quantitatively detecting KRECs gene and application thereof. The kit comprises a PCR system based on nested PCR technology and a real-time fluorescence quantitative PCR system based on real-time fluorescence PCR technology, wherein the nested PCR system comprises forward and reverse primers for KRECs and beta-actin genes; and the real-time fluorescence quantitative PCR system comprises forward and reverse primers and specific fluorescence probes for KRECs and beta-actin genes. The kit can be used for quickly screening the B-cell level of a neonatal immune system, and has the advantages of high sensitivity, high stability and excellent reproducibility. The method is suitable for quantitative detection of KRECs, can be used for screening functions of the neonatal immune system, and has practical clinical application value.

The invention relates to a real-time fluorescence quantitative PCR (polymerase chain reaction) kit for quantitatively detecting TRECs gene and application thereof. The kit comprises a PCR system based on nested PCR technology and a real-time fluorescence quantitative PCR system based on real-time fluorescence PCR technology, wherein the nested PCR system comprises forward and reverse primers for TRECs and beta-actin genes; and the real-time fluorescence quantitative PCR system comprises forward and reverse primers and specific fluorescence probes for TRECs and beta-actin genes. The kit can be used for quickly screening the T-cell level of a neonatal immune system, and has the advantages of high sensitivity, high stability and excellent reproducibility. The method is suitable for quantitative detection of TRECs, can be used for screening functions of the neonatal immune system, and has practical clinical application value.

The invention belongs to the field of biological detection, and particularly relates to a detecting method for carrying out single-tube ternary reverse transcription on three kinds of miRNA including U6, miR 92a and miR 21, a detecting method for carrying out single-tube double quantification respectively after reverse transcription and a kit. The kit comprises a reverse transcription stem-loop primer group, a quantitative detecting primer group, a quantitative detecting probe group, an enzyme system and a PCR reaction reagent; reverse transcription of U6, miR 92a and miR 21 can be carried out in the same reaction tube; meanwhile, by high-sensitivity and high-specificity quantitative detecting primer sequences and quantitative detecting probes and PCR reaction procedures and conditions which are matched with the high-sensitivity and high-specificity quantitative detecting primer sequences and quantitative detecting probes, miR 21 and U6 as well as miR 92a and U6 can be subjected to single-tube double quantification, operation errors during single-tube single quantification in the prior art are avoided, and quantification is simple and accurate. In addition, a double-polymerase amplification for Stoffel fragments and Tfl DNA polymerase is introduced, while detection specificity is improved, lots of templates can be tolerated, and detection sensitivity and result stability are improved.

The invention belongs to the technical field of molecular biology, and in particular relates to a human platelet antigen genotyping liquid phase chip and a detection method thereof. Human platelet antigen genotyping technology is an important technical means to provide patients with matching platelets and promote the safety and effectiveness of clinical platelet transfusion. However, the existing human platelet antigen genotyping technology has low throughput and high accuracy. Indexes such as sensitivity and sensitivity need to be improved. The liquid phase chip of the present invention includes primers 1-24, connection probes 25-46 and fluorescently encoded microspheres coated with detection probes 47-68, and the human platelet antigen gene is completed through amplification, connection and hybridization reactions. Typing detection. The liquid phase chip and detection method adopted in the present invention have the advantages of large flux, high sensitivity, strong specificity, good repeatability, wide linear range, simple operation, strong flexibility, and wide application range, and are clinically accurate, Efficient and practical human platelet antigen genotyping detection method.

The invention relates to an oligonucleotide primer probe for precise quantitative detection of specific gene components of the transgenic rice Kefeng No. 6 strain, and a kit comprising the primer probe. The present invention also relates to a digital PCR detection method for quantitatively detecting the specific gene components of the transgenic rice Kefeng No. 6 strain, said method comprising using specific oligonucleotide primers and Fluorescently labeled probes. The invention also relates to the application of the specific oligonucleotide primers and fluorescent probes for the specific genes of the transgenic rice strains in the quantitative detection of the specific gene components of the transgenic rice Kefeng No. 6 strain. The digital PCR detection method of the present invention can accurately and sensitively measure the content of specific gene components of the transgenic rice Kefeng No. 6 strain in the sample, and the sensitivity can reach 1 copy / μL.

The invention provides a quantitative analysis method for integrating proteome and glycoproteome, which comprises the following steps: (1) performing reductive alkylation treatment after heating and denaturing the protein; (2) using trypsin in an ultrafiltration tube to convert the protein Cut peptides with enzymes and centrifuge to obtain peptides; (3) Hydrophilic interaction chromatography HILIC filler binds peptides, washes and removes impurities, and then elutes to obtain complete N-glycopeptides; (4) Adds trifluoroacetic acid and heats to remove N-glycopeptides ‑Sialic acid on the periphery of the sugar chain to obtain asialic N‑glycopeptide; (5) Use HCD‑MS / MS to analyze the peptide to obtain proteome data, and use MaxQuant for qualitative and quantitative analysis; (6) Use SCE‑HCD‑MS / MS analysis of intact N-glycopeptides and asialo-N-glycopeptides, using Xcalibur software for qualitative and quantitative analysis. The invention can simply, quickly and effectively realize the quantitative analysis of proteome and glycoproteome, and has a good application prospect in the study of the occurrence and development mechanism of diseases and the discovery of new biomarkers.

The invention relates to a real-time fluorescence quantitative PCR (polymerase chain reaction) kit for quantitatively detecting TRECs and KRECs genes and application thereof. The kit comprises a PCR system based on nested PCR technology and a real-time fluorescence quantitative PCR system based on real-time fluorescence PCR technology, wherein the nested PCR system comprises forward and reverse primers for TRECs, KRECs and beta-actin genes; and the real-time fluorescence quantitative PCR system comprises forward and reverse primers and specific fluorescence probes for TRECs, KRECs and beta-actin genes. The kit can be used for quickly screening the T-cell level and B-cell level of a neonatal immune system, and has the advantages of high sensitivity, high stability and excellent reproducibility. The method is suitable for combined quantitative detection of TRECs and KRECs, can be used for screening functions of the neonatal immune system, and has practical clinical application value.

The invention relates to a water body pollutant detection method, in particular to a rapid detection method for imidacloprid water body pollution by fluorescent quantitative PCR, belonging to the field of molecular biology. The present invention detects the mitochondrial gene of the aquatic insect Yixing Kuanjima wxya The technology of imidacloprid pollution monitoring based on the response of expression level to imidacloprid, the specific implementation steps are as follows: (1) cultivation of Yixing broad-based mayfia in water samples to be tested; (2) extraction of total RNA; (3) gel electrophoresis detection ; (4) Measurement of RNA concentration; (5) Inversion of RNA into cDNA; (6) Construction of standard products and creation of standard curve; (7) Measurement of gene expression; (8) Data analysis. The method has high sensitivity, good timeliness, strong specificity, simple operation, and can quickly detect whether the water body is polluted by imidacloprid.

The disclosure belongs to the field of genome editing efficiency detection, and specifically relates to a genome editing detection method, kit and application. In view of the defects of gene editing efficiency detection methods in the prior art, such as Sanger, NGS, and methods based on mismatch-specific nucleases, etc. have defects such as complicated operation, high cost, and insufficient detection accuracy, the present disclosure provides a new method , called getPCR, based on Taq DNA polymerase specificity and real-time PCR technology, quantifies the wild-type DNA in the genome to be tested, and confirms the genome editing efficiency by calculating the percentage of wild-type DNA. This disclosed research provides the design rules of the corresponding guarded bases and optimized getPCR operation parameters. It has been verified that the method has good detection accuracy and is easy to operate. It can be applied to all genome editing methods to quantify genome editing efficiency, and can also be applied to Screening of single cell clones.

The invention discloses a paper-based double-antibody sandwich method based on a covalent bonding method to fix and capture antibodies, belongs to the technical field of immune analysis, and is used for detecting the content of allergy-like related MRGPRX2 receptors in human blood. Using MRGPRX2 mouse monoclonal antibody as the capture antibody, without the need to chemically modify the surface of the filter paper, by introducing bovine serum albumin (BSA) molecules with both carboxyl and amino functional groups, using the amide condensation reaction between the antibody molecule and the amino and carboxyl groups in the BSA molecule The antibody-BSA network structure was formed to realize the immobilization of the capture antibody; the MRGPRX2 rabbit polyclonal antibody labeled with horseradish peroxidase HRP was used as the detection antibody, and the MRGPRX2 solution was used as the standard to establish a paper-based double-antibody sandwich method for MRGPRX2. The immobilization method of the capture antibody is simple, efficient, and low in cost. The technical indicators of the detection method are high in detection throughput, fast in speed, detection limit and precision, etc., and the accuracy and reproducibility of the quantitative results are good. It is suitable for For clinical examination and blood epidemiological investigation.

The invention provides a quantitative analysis method of liquid-state products of a hydrocarbon generation and expulsion simulation experiment. The method comprises the following steps: collecting and treating the liquid-state products of the hydrocarbon generation and expulsion simulation experiment, wherein the step comprises light hydrocarbon collecting, heavy hydrocarbon collecting, standard sample adding, dehydrating, filtering and concentrating; carrying out quantitative analysis on the liquid-state products of the hydrocarbon generation and expulsion simulation experiment by utilizing a comprehensive two-dimensional gas chromatography-hydrogen flame ionization detector, wherein the step comprises primary comprehensive two-dimensional analysis, natural volatilization for constant weight as well as secondary comprehensive two-dimensional analysis; calculating full-component quantitative result of the liquid-state products of the hydrocarbon generation and expulsion simulation experiment. The quantitative analysis method disclosed by the invention can be used for avoiding volatilization of light hydrocarbon, has better experiment result repeatability and simple and easy-to-learn operation, and provides a reliable technical method for the quantitative analysis of the liquid-state products of the hydrocarbon generation and expulsion simulation experiment, so that estimation on basin oil and gas resource amount is more objective.

The invention belongs to the field of biological medicine, relates to a specific high-affinity recombinant antibody marking fluorescent microsphere and a preparation method and application thereof and particularly relates to a plug-and-play fluorescence detection card for quick quantitative detection of KIM-1, NGAL, IGFBP7, TIMP-2, IL-18, PCT and cTnI in human urine or blood samples. The method includes: activating fluorescent microspheres; adding the activated fluorescent microspheres into a dialysis bag filled with phosphate buffer to obtain dialyzed fluorescent microsphere solution; adding amino-polyethylene glycol-carboxyl, further activating the fluorescent microsphere solution subjected to secondary dialysis, making with a recombinant antibody, centrifuging after reaction, washing, precipitating, resuspending and precipitating. The fluorescent microsphere has advantages that immunofluorescence reaction lower limit can be remarkably improved to enable sensitivity of a prepared test card to be improved by 10-100 times of that of test cards prepared according to a traditional method, and a test range is remarkably expanded.