54 results about "Denaturing high performance liquid chromatography" patented technology

The present invention relates to a cancer biomarker molecule, a screening test kit and a detection method thereof. According to the present invention, analysis of methylating areas of specimen target genes PAX1, ZNF582, SOX1 and NKX6-1 are adopted, plural of segments of target gene-specific oligonucleotide primers or probes are designed, and the oligonucleotide probes are adopted to detect presence or absence of methylation of the target genes so as to further determine possibility of cancer occurrence; and the methylation state detection method comprises methylation-specific PCR (MSP), quantitative methylation-specific PCR (QMSP), bisulfite sequencing (BS), microarrays, mass spectrometer, denaturing high-performance liquid chromatography (DHPLC), pyrosequencing or Enzyme-linked immunosorbent assay (ELISA) and the like.

The invention provides a group of nucleic acids used in a quintuple multiplex polymerase chain reaction (mPCR)-denaturing high-performance liquid chromatography (DHPLC) method for detecting mycobacterium and identifying pathogenic mycobacterium. The nucleic acids comprise five pairs of primers of which the nucleic acid sequences are shown as SEQ ID No.1 and SEQ ID No.2, SEQ ID No.4 and SEQ ID No.5, SEQ ID No.7 and SEQ ID No.8, SEQ ID No.10 and SEQ ID No.11, and SEQ ID No.13 and SEQ ID No.14, and PCR amplification products which are used as positive control and of which the nucleic acid sequences are shown as SEQ ID No.3, SEQ ID No.5, SEQ ID No.9, SEQ ID No.12 and SEQ ID No.15. The invention also provides a kit using the nucleic acids and a detection method; the method is high in specificity and flexibility and easy to operate, and high flux can be achieved; and the method has important practical significance to clinical identification on the mycobacterium infection and infectious agents of the mycobacterium infection.

The invention discloses a method for extracting DNA from old formalin-fixed tissues. The basic principle of the method is to utilize the microwave thermal effect for fast removing formalin in the fixed tissues via water molecules, and eliminate the DNA-protein crosslinking caused by aldehyde groups of the formalin, thereby completely or partially recovering a DNA structure and further obtaining the DNA with better quality. The invention relates to the simple and high-efficient method for extracting the DNA from the old formalin-fixed tissues, which is based on microwave thermal remediation and can obtain the DNA with higher quality (large fragments and high purity), so that the method can be used for multiplex fluorescent PCR-capillary electrophoresis (FM-CE), denaturing high performance liquid chromatography (DHPLC) analysis, direct DNA sequencing and other molecular biological method analyses.

The invention discloses a food foodborne pathogenic bacteria rapidly identifying method. The food foodborne pathogenic bacteria rapidly identifying method comprises the steps of 1) DNA (deoxyribonucleic acid) template extraction, 2) primer screening and reaction system optimization including selecting the specific genes of nuc, invA and prfA of staphylococcus aureus, salmonellas and listeria monocytogenes to separately design primer pairs, 3) PCR (polymerase chain reaction) amplification, 4) PCR-DHPLC (polymerase chain reaction-denaturing high performance liquid chromatography) detection of strain DNA, during which only strains coinciding with target amplification can be detected with amplification absorption peaks while other non-targeted amplification bacteria detected to be negative. The food foodborne pathogenic bacteria rapidly identifying method can detect three foodborne pathogenic bacteria in food when applied every single time, thereby saving time and consumable agent, simplifying operation, avoiding excessive artificial errors and achieving high sensitivity.

The invention discloses a PCR-DHPLC (polymerase chain reaction-denaturing high performance liquid chromatography) assay primer and assay method for transgenic rapeseed RF2 strain. The primer is strong in specificity, and can be used in PCR and used for specifically amplifying a DNA (deoxyribonucleic acid) fragment of the transgenic rapeseed RF2 strain, and the amplified product can be applied to subsequent DHPLC analysis. The assay method provides an assay method for the transgenic rapeseed RF2 strain, which is convenient to operate, good in extension performance and strong in specificity. The PCR amplified product is analyzed by utilizing the DHPLC, the resolution of the fragment of the PCR amplified can be a plurality of basic groups, and the resolution rate is high. The primer and assay method provided by the invention provide a simple, convenient, effective and reliable assay method to assay of the transgenic rapeseed RF2 strain, and are particularly applicable to port inspection and quarantine and other departments.

The invention discloses a primer and a kit for detecting methylation of MGMT (O<6>-methylguanine-DNA methyltransferase) genes, a use method of the primer and application of the primer and the kit. The primer disclosed by the invention is a specific primer designed according to sequences of promoter regions of the human MGMT genes, and the kit containing the primer can be used for rapidly and accurately detecting the methylation of the MGMT gene with high sensitivity through the combination of nest-shaped PCR (Polymerase Chain Reaction) amplification and DHPLC (Denaturing High Performance Liquid Chromatography) detection.

Methods, compositions, and kits for separating heteroduplex and homoduplex DNA molecules in a test mixture by temperature-compression denaturing high performance liquid chromatography (tcDHPLC). The method includes use of nitrogen-containing additives in the mobile phase that allow detection of diverse heteroduplex molecules to be performed at the same pre-selected temperature. An example of a preferred additive is betaine. Standard mixtures of DNA fragments, such as mutation standards containing known heteroduplex and homoduplex molecules, can be used to select the concentration of additive and temperature. Compositions and kits including the mobile phase, mutation standards, PCR primers, separation media, and DNA polymerase are also provided.

The invention discloses a PCR-DHPLC (polymerase chain reaction-denaturing high performance liquid chromatography) detection primer and a detection method for genetically modified rice strain KMD. The primer has strong specificity and can be used for PCR amplification and DHPLC analysis. The detection method is simple and convenient to operate, good in expansion performance and high in sensitivity. The DHPLC is used to analyze PCR amplified products, fragment size resolution can reach multiple bases, and resolution ratio is high. The PCR-DHPLC detection primer and the detection method for genetically modified rice strain KMD have the advantages that the detection method is simple, convenient, effective, reliable and especially suitable for departments of port inspection and quarantine and the like.

The invention discloses a multiplex PCR-DHPLC (polymerase chain reaction-denaturing high performance liquid chromatography) detection primer and a detection method for genetically modified maize. The primer has strong specificity and can be used for PCR amplification and DHPLC analysis. The detection method is simple and convenient to operate, good in expansion performance and high in sensitivity, and multi-target detection of the genetically modified maize is realized. The DHPLC is used to analyze PCR amplified products, fragment size resolution can reach multiple bases, and resolution ratio is high. The multiplex PCR-DHPLC detection primer and the detection method for genetically modified maize have the advantages that the detection method is simple, convenient, effective, reliable, high in throughput and especially suitable for departments of port inspection and quarantine and the like.

The invention discloses a PCR-DHPLC (polymerase chain reaction-denaturing high performance liquid chromatography) assay primer and assay method of transgenic rapeseed RT73 strain. The primer is strong in specificity, and can be used in PCR and used for specifically amplifying a DNA (deoxyribonucleic acid) fragment of the transgenic rapeseed RT73 strain, and the amplified product can be applied to subsequent DHPLC analysis. The assay method provides an assay method for the transgenic rapeseed RF2 strain, which is convenient to operate, good in extension performance and strong in specificity. The PCR amplified product is analyzed by utilizing the DHPLC, the resolution of the fragment of the PCR amplified can be a plurality of basic groups, and the resolution rate is high. The primer and assay method provided by the invention provide a simple, convenient, effective and reliable assay method to assay of the transgenic rapeseed RT73 strain, and are particularly applicable to port inspection and quarantine and other departments.

The invention relates to specific primers for detecting swine infuenza virus subtype H5 by using DHPLC (denaturing high performance liquid chromatography) and application thereof. The specific primers are nucleotide sequences disclosed as SEQ ID NO.1 and SEQ ID NO.2. By adopting the PCR (polymerase chain reaction) amplification-nondenatured DHPLC combined analysis mode, the invention establishes a new method for quickly screening swine infuenza viruses. The method can quickly and automatically screen swine infuenza viruses at high flux, greatly enhances the detection efficiency, shortens the detection time, and has important clinical meanings for preventing and controlling outbreak of swine infuenza subtype H5.

The invention discloses fasciolopsis PCR-DHPLC (polymerase chain reaction-denaturing high performance liquid chromatography) detection primers, a kit and a detection method. Primers SEQ ID NO.1-2 are designed according to the gene sequence of fasciolopsis, and a PCR-DHPLC method is utilized to carry out qualitative detection on the fasciolopsis in food or aquatic plants. The kit comprises a 5U/mu L Taq DNA (deoxyribonucleic acid) polymerase, 2.5 mM of PCR reaction solution, 2.5 mM of dNTP (deoxyribonucleotide triphosphate), 10 mu M of SEQ ID NO.1 and 10 mu M of SEQ ID NO.2. The method can detect the fasciolopsis to the precision of 1 fasciolopsis/gram sample. The invention has the advantages of short time consumption and simple operation, can save abundant human resources and material resources, and is suitable for the requirements of quick detection.

The invention relates to a soil microbial diversity analysis method, belonging to the technical field of soil microbial analysis. In order to solve the problems of heavy pollution and long time consumption, the soil microbial diversity analysis method is provided. The soil microbial diversity analysis method includes microbial genome DNA extraction of acquired soil to obtain a genome DNA extract; detection of the genome DNA extract to determine the soil genomic DNA concentration and purity; PCR amplification of DNA extracted from the soil to obtain a PCR amplification product; and microbial diversity analysis of the PCR amplification product by use of denaturing high Performance liquid chromatography. According to the method, primer marking is not needed, glue leaking using toxic substance formamide and acrylamide is not needed, the radioactive pollution is reduced, and the soil microbial diversity analysis method has the effects of less pollution, high degree of automation, short detection time, high sensitivity, and good detection strip separation effect.

The invention relates to a detection kit for Campylobacter jejuni in pork. The detection kit comprises a culture medium and a detection solution, wherein the culture medium is broth. The detection solution is composed of 3-5 mu l of 10 PCR (polymerase chain reaction) buffer solution, 0.2-0.4 mu l of Taq DNA (deoxyribonucleic acid) polymerase, 3-5 mu l of dNTP (deoxyribonucleotide triphosphate), 0.1-0.6 mu l of 100bp DNA Ladder Marker, 1-2 mu l of PCR primer, 5-10 ml of Tris.cl, 1-2.2mg of sodium chloride, 1-2.2mg of magnesium chloride, 0.2-0.5mg of disodium hydrogen phosphate dodecahydrate and 3-5mg of enrichment broth. The detection kit is an improvement on the basis of the conventional PCR technology, and can be used for qualitative detection of Campylobacter jejuni. By integrating the advantages of PCR, DHPLC (denaturing high performance liquid chromatography) and other techniques, the detection kit has the advantages of high specificity of detection results and high sensitivity, and ensures the effective detection of Campylobacter jejuni in food.

The invention discloses a trichina PCR-DHPLC (Polymerase Chain Reaction-Denaturing High Performance Liquid Chromatography) detection primer as well as a kit and a detection method. In allusion to a gene sequence of the trichina, detection primers SEQ ID NO.1-2 are designed and a PCR-DHPLC method is used for the qualitative detection of the trichina in detected animal derived food. The kit comprises Taq DNA polymerase with concentration of 5U/microlitre, 2.5mM of PCR reaction liquid, 2.5mM of dNTP (Diethyl-Nitrophenyl Thiophosphate), 10 microns of SEQ ID NO.1 and 10 microns of SEQ ID NO.2. The method disclosed by the invention can be used for precisely detecting the polypide sample of one trichina, and has the advantages of being short in detection time and simple to operate, saving lots of manpower and material resources and being suitable for rapid detection.

The invention discloses a PCR-DHPLC (polymerase chain reaction-denaturing high performance liquid chromatography) detection primer and a detection method for genetically modified maize strain BT11. The primer has strong specificity and can be used for PCR amplification and DHPLC analysis. The detection method is simple and convenient to operate, good in expansion performance and high in sensitivity. The DHPLC is used to analyze PCR amplified products, fragment size resolution can reach multiple bases, and resolution ratio is high. The PCR-DHPLC detection primer and the detection method for genetically modified maize strain BT11 have the advantages that the detection method is simple, convenient, effective, reliable and especially suitable for departments of port inspection and quarantine and the like.

The invention discloses a primer and a method for PCR-DHPLC (polymerase chain reaction-denaturing high-performance liquid chromatography) detection for endogenous genes of wheat. The primer is high in specificity and can be used for PCR amplification and DHPLC analysis. The method for detection for the endogenous genes of the wheat is simple in operation, good in extensibility and high in sensitivity. PCR amplification products are analyzed by means of DHPLC, the segment size resolution of the PCR amplification products can reach multiple basic groups, and the resolution ratio is high. The primer and the method have the advantages that the method is a simple, convenient, effective and reliable method for detection for the endogenous genes of the wheat, and the primer and the method are particularly suitable for port inspection and quarantine departments and the like.

The invention discloses a PCR-DHPLC (polymerase chain reaction-denaturing high performance liquid chromatography) assay primer and assay method for transgenic rapeseed MS1 strain. The primer is strong in specificity, and can be used in PCR and used for specifically amplifying a DNA (deoxyribonucleic acid) fragment of the transgenic rapeseed MS1 strain, and the amplified product can be applied to subsequent DHPLC analysis. The assay method provides an assay method for the transgenic rapeseed MS1 strain, which is convenient to operate, good in extension performance and strong in specificity. The PCR amplified product is analyzed by utilizing the DHPLC, the resolution of the fragment of the PCR amplified can be a plurality of basic groups, and the resolution rate is high. The primer and assay method provided by the invention provide a simple, convenient, effective and reliable assay method to assay of the transgenic rapeseed MS1 strain, and are particularly applicable to port inspection and quarantine and other departments.