12 results about "High Resolution Melt Analysis" patented technology

The invention belongs to the technical field of traditional Chinese medicine identification, and particularly relates to a primer pair for identifying honeysuckle and lonicerae flos and application of the primer pair. The sequences of the primer pair are as shown in SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 9 and SEQ ID NO: 10. According to the present invention, after the PCR amplification is performed on the honeysuckle and the lonicerae flos by using the primer pair, the high resolution melting curve and the gel electrophoresis band of the PCR amplification product are analyzed to discover that the typing effect is good, and the honeysuckle and lonicerae flos samples with different mass ratios can be distinguished so as to achieve the identification of the honeysuckle and the lonicerae flos.

Methods of determining methylation of DNA are provided that include filtering a biological sample comprising a nucleic acid with a first matrix material to purify the DNA; eluting and denaturing the DNA to produce eluted denatured DNA; heating the DNA in the presence of bisulfite ions to produce deaminated nucleic acid; optionally contacting said deaminated nucleic acid with a second matrix material; desulphonating and eluting the bound deaminated nucleic acid; and performing methylation specific PCR, nucleic acid sequencing, and / or high resolution melting analysis (HRM) on said bisulfite-converted nucleic acid to determine the methylation of said nucleic acid, wherein multiple steps may be performed in a single reaction cartridge.

The invention relates to the technical field of biomedical detection, and discloses a kit product for detecting SNP (Single Nucleotide Polymorphism) of human micro ribonucleic acid absorption capacity, which comprises one or more of an SNP1 site and an SNP2 site. The method has the advantages that the operation process is simple, and the capacity of the main body of the detected sample for absorbing the micro ribonucleic acid can be determined according to the condition of the SNP site only through the two steps of DNA extraction and high-resolution melting curve method PCR reaction; the detection sample is more stable and easier to store; the method can provide help for drug effect identification of drugs containing micro ribonucleic acid and determination of clinical medication of the drugs, can improve the detection efficiency, and has a wide application prospect.

The invention discloses a human MTHFR (methylenetetrahydrofolate reductase), MTRR (methylenetetrahydrofolate reductase) and SLC19A1 gene detection kit. The kit is based on a high-resolution melting curve analysis technology, and comprises four groups of primer pairs aiming at C677T and A1298C sites of MTHFR genes, A66G sites of MTRR genes and A80G sites of SLC19A1 genes. The kit can be used for rapidly, sensitively and accurately identifying the genotypes of the four key sites under the same reaction procedure by detecting the melting temperature of PCR products and the difference of curve shapes. The kit is suitable for risk assessment of cardiovascular diseases and birth defects, is more innovatively applied to donor screening and receptor assessment of coprophilous fungus transplantation, fills the blank of genetic background detection tools in the field, and has the advantages of low cost, high sensitivity, high specificity, simplicity and convenience in operation, high repeatability, quick and objective detection result and the like.

The present application belongs to the cross field of molecular diagnosis, environmental microorganism detection and nanotechnology, and particularly relates to a biological aerosol automatic detection platform, a detection method and application. The present application realizes specific capture and enrichment of target microorganisms through a silicon nanotip array, and the silicon nanotip array chip is provided with a plurality of regularly arranged silicon nanotips, and the surface of the silicon nanotip is modified with biological capture molecules. The temperature change of the nanotip is monitored in real time by using Raman spectrum, and the feedback optimization of the lysis / amplification temperature curve is realized, the double signal amplification and verification of LAMP amplification and CRISPR detection are combined, the high-resolution melting curve is analyzed through a machine learning algorithm, and the automatic identification and molecular typing of the strain are realized. The present application has the advantages of high detection sensitivity, strong specificity, simple operation, short detection time and the like. The present application is suitable for on-site rapid detection and real-time monitoring of pathogenic bacteria in various environments such as atmosphere, exhaled air and water.

The invention discloses a specific molecular marker for identifying genetic sex of mugilogobius chuyi and application of the specific molecular marker for identifying genetic sex of mugilogobius chuyi. According to the invention, a specific molecular marker capable of identifying genetic sex of mullet chubil is developed by sequencing, comparing and analyzing full-length transcriptome of male and female gonads, and a detection primer pair of the molecular marker is designed to carry out PCR and qPCR amplification detection on a to-be-detected mullet chubil sample. Through Sanger sequencing and peak patterns of a high-resolution melting curve, the genetic sex of the mullet chubil can be accurately distinguished, and verification shows that the method is stable and applicable to both laboratory groups and wild groups. The method for rapidly, accurately, economically and reliably identifying the genetic sex of the mullet obliterans is established, is suitable for identifying the genetic sex of the mullet obliterans in each development stage, can conveniently and rapidly confirm the genetic sex of the mullet obliterans, and lays a foundation for researching the biological characteristics of the mullet obliterans and sex differentiation and regulation of the mullet obliterans.

The invention discloses a method for improving screening accuracy of specific primers of Shigella by using fluorescent loop-mediated isothermal amplification and a high-resolution melting curve, and belongs to the field of molecular biology. The method comprises the following steps: designing a primer group through an ipaH gene, extracting Shigella genome DNA, taking the Shigella genome DNA as a template, carrying out a fluorescent LAMP amplification reaction on the designed LAMP primer group, and carrying out HRM analysis on an amplification product; the positive amplification curve in the amplification curves is in a typical S shape, the negative control is not amplified, or the peak time of the amplification curve lags the positive peak time by more than 30 min is screened as a specific primer; in an HRM melting curve, a positive template is in a single sharp peak shape, and the positive Tm value is 90.0-91.0 DEG C; the negative control does not have a specific melting peak or has a wide and blunt non-specific peak, and the difference value between the negative control Tm and the positive Tm is greater than or equal to 2.0 DEG C. Through dual verification of fluorescent LAMP and HRM accurate analysis, the advantages and disadvantages of the primers are accurately evaluated, accurate and efficient screening of the optimal primer group is realized, and sensitive and specific primers are provided for subsequent LAMP detection tests.

The present invention relates to a method for detecting and identifying a microorganism in a biological sample by a nucleic acid amplification reaction, said method comprising the steps of: providing a biological sample suspected of comprising a microorganism, and optionally isolating nucleic acids from said biological sample; b) providing a reaction container comprising a nucleic acid amplification reaction mixture comprising an aliquot of said biological sample or nucleic acids isolated therefrom, at least one pair of broad-taxonomic-range amplification primers for amplifying a microbial ribosomal DNA (rDNA) internal transcribed spacer (ITS) region from the, optionally isolated, nucleic acids in said sample and for generating an rDNA ITS amplicon, at least one taxon-specific detection probe for detecting a taxon-specific sequence in said rDNA ITS region, a DNA intercalating dye for recording a high resolution melting curve for said rDNA ITS amplicon; performing a nucleic acid amplification reaction on the mixture to generate said rDNA ITS amplicon; recording an hrMC for said rDNA ITS amplicon; comparing the high resolution melting curve recorded with a database comprising hrMCs of reference amplicons generated from reference microbial species of known taxonomic identity using the same set of broad-taxonomic-range amplification primers, to thereby obtain a first taxonomic identity indicator of a microorganism present in said sample; detecting hybridization between said at least one taxon-specific detection probe and said rDNA ITS amplicon to thereby obtain a second taxonomic identity indicator of a microorganism present in said sample, and identifying the microorganism present in said sample at species level based on said first and second taxonomic identity indicator, wherein steps c), d), and f) are carried out in a closed reaction container.

The invention provides a fine-wool sheep wool quality character related gene detection kit and method, and belongs to the technical field of animal molecular breeding. The kit is based on 15 genetic variation sites of eight key functional genes such as KRTAPs, FGF5 and BMP2, target fragments are amplified through multiple PCR, genetic typing is achieved by combining high-resolution melting curve analysis, Sanger sequencing or CRISPR / Cas12a lateral flow detection, and wool quality genetic indexes are constructed for comprehensive evaluation. The kit adopts preloaded freeze-dried microspheres and partitioned sealing, so that the transportation stability and the operation convenience are ensured. The method can complete detection in the early stage of lambs, significantly shortens the breeding cycle, and improves the accuracy of seed selection. The method has the advantages of multi-gene joint detection, flow standardization, result visualization, wide application range and the like, and is suitable for genetic improvement and efficient breeding popularization of fine-wool sheep.

Methods of determining methylation of DNA are provided that include filtering a biological sample comprising a nucleic acid with a first matrix material to purify the DNA; eluting and denaturing the DNA to produce eluted denatured DNA; heating the DNA in the presence of bisulfite ions to produce deaminated nucleic acid; optionally contacting said deaminated nucleic acid with a second matrix material; desulphonating and eluting the bound deaminated nucleic acid; and performing methylation specific PCR, nucleic acid sequencing, and / or high resolution melting analysis (HRM) on said bisulfite-converted nucleic acid to determine the methylation of said nucleic acid, wherein multiple steps may be performed in a single reaction cartridge.

The invention relates to a marker for detecting microsatellite instability of endometrial cancer, a kit and application. The marker composition comprises a first marker, a second marker, a third marker, a fourth marker and a fifth marker. The marker composition which is higher in sensitivity, better in specificity and stronger in compatibility to the endometrial cancer microsatellite instability is obtained through screening, when the marker composition is used for detecting the endometrial cancer microsatellite instability under the condition that only five markers (HAL, NBAS, ZBTB37, TSHZ2 and FOXP2) are contained, the coincidence rate of the detection result and an IHC method is as high as 100%, and the marker composition can be used for detecting the endometrial cancer microsatellite instability. According to the present invention, the endometrial cancer detection performance is far higher than the endometrial cancer detection performance of the similar high-resolution melting curve method site composition reported at present, such that the detection cost can be effectively reduced, the efficiency can be effectively improved, the detection lower limit can achieve the tumor DNA content of 5%, the result bimodal map distinguishing is significant, the result determination is convenient, and the overall detection performance is excellent.