9 results about "Dna integrity" patented technology

The application discloses a frozen embedding block of islet cell-like cell clusters meeting spatiotemporal omics continuous section analysis, a method and application thereof, and belongs to the field of frozen embedding block preparation. In order to solve the problem of RNA / DNA integrity and visualization of islet cell clusters, the method comprises the following steps: adding a buffer solution containing bromophenol blue drop by drop to the aggregated cell clusters on the inner surface of the bottom of a silk cloth to dye the cell clusters; keeping the selected area of the silk cloth in contact with the frozen embedding reagent surface of the lower half groove in the loading groove of a first mold, placing the first mold on dry ice, and completely freezing the frozen embedding reagent of the lower half groove and the cell clusters on the surface thereof; placing the first mold with the upper half groove frozen embedding reagent added in the loading groove on dry ice, completely freezing the frozen embedding reagent of the upper half groove and the cell clusters on the surface thereof, and making the frozen embedding reagent of the upper half groove and the frozen embedding reagent of the lower half groove be integrated into the frozen embedding block, and taking out the frozen embedding block, wherein the cell clusters are distributed at the middle position on the height of the frozen embedding reagent block.

This invention discloses a plasmon resonance detection method for hepatobiliary-specific methylation sites. The method first immobilizes a sequence-specific DNA capture probe on a gold membrane surface and covalently couples it with a methylation-binding domain protein, constructing a sensor interface with dual functions of sequence capture and methylation recognition. During detection, a pre-treated plasma circulating cell-free DNA single-stranded sample is introduced into the sensor. The target DNA first hybridizes with the capture probe, and then its methylation site is specifically bound by the adjacent methylation-binding domain protein, triggering two consecutive specific surface plasmon resonance signal responses. By analyzing the increment between the two stable signal values ​​and comparing it with a preset threshold, label-free, real-time, and highly specific determination of the methylation status of the target site can be achieved. This invention eliminates the need for bisulfate treatment, effectively preserving DNA integrity, and is particularly suitable for high-sensitivity detection of hepatobiliary disease-related epigenetic markers in trace circulating cell-free DNA.

The application provides an On-DNA 2,5-disubstituted thiophene leading compound and a synthesis method thereof, and belongs to the field of gene coded compound library construction. The method has small DNA damage, good universality, simple operation, mild conditions and can obtain the On-DNA 2,5-disubstituted thiophene compound with high conversion rate. Under the reaction conditions in the application, the product On-DNA 2,5-disubstituted thiophene compound not only has high conversion rate, but also has good DNA integrity. The application enriches the chemical reaction type of the synthesis of the coded compound library on the DNA, constructs a new 2,5-disubstituted thiophene skeleton for the gene coded compound library, and has very good application prospect in the development of leading drugs.

This application provides a method and related equipment for DNA integrity assessment based on single sperm Raman spectroscopy, belonging to the field of sperm quality assessment technology in assisted reproductive technology. This method, based on a machine learning algorithm, directly assesses DNA integrity using the Raman spectra of single sperm, thereby achieving non-destructive screening of individual high-quality sperm in assisted reproductive technology. The process of constructing the algorithm training sample dataset includes a step-by-step fixation and analysis method for sample single sperm: first, Raman spectroscopy testing is performed, followed by fluorescence staining analysis, reducing the impact of sperm position changes during detection, analysis, or staining. Then, optical and fluorescence images of single sperm within preset shape units are matched to determine the validity of the sample single sperm. Finally, a training sample dataset is constructed based on the relevant data of valid single sperm, establishing a correlation between the Raman spectrum of the same single sperm and its sperm chromatin structure analysis results.

The invention discloses an endangered species-oriented RAD-seq low-cost genetic diversity evaluation system, and relates to the technical field of biology, the endangered species-oriented RAD-seq low-cost genetic diversity evaluation system comprises an evaluation system, the evaluation system comprises a DNA extraction and preprocessing module, a simplified RAD-seq library building module, a high-throughput sequencing module and a genetic diversity analysis module, the DNA extraction and pretreatment module is used for extracting low-quality DNA from a non-invasive sample of an endangered species and carrying out DNA repair and concentration standardization, the simplified RAD-seq library building module adopts a low-cost self-made reagent combination to replace a commercial kit, and by using the self-made reagent combination and simplifying a library building process, such as a single-step enzyme digestion-ligation reaction, the library building efficiency is greatly improved, and the application range of the library building module is widened. The method has the advantages that the method is simple, the reagent and operation cost is obviously reduced, the library building time is shortened, the method is suitable for large-scale endangered species research, the DNA integrity and the library building success rate are improved by performing end repair and notch filling on degraded DNA through a repair unit of the DNA extraction and pretreatment module, and the method is particularly suitable for non-invasive samples.

The present application relates to the field of biotechnology and molecular diagnosis, and particularly relates to a methylation detection pretreatment kit and method for protecting DNA integrity, which can solve the problem that DNA degradation is serious in traditional bisulfite conversion technology, and conversion efficiency and DNA integrity are difficult to be obtained simultaneously, the kit comprises a conversion solution, a protection solution, a combination solution, a desulfonation solution, a washing solution and an elution solution, the conversion solution contains 2.5-4 mol / L bisulfite and linear polyethylene polyamine, the protection solution contains an organic denaturant and a polyphenol antioxidant, the DNA melting temperature is reduced by the organic denaturant, the linear polyethylene polyamine accelerates the conversion reaction, and the desulfonation system with low water activity is combined to complete the bisulfite conversion under mild conditions of 50-70 DEG C; the present application can realize the conversion efficiency of greater than 99.5% of unmethylated cytosine, greatly reduce DNA degradation, significantly improve the long fragment DNA recovery rate and the detection sensitivity of trace samples, and is suitable for the methylation detection of three generations of long read sequencing and clinical trace samples.

A retrotransposable element based multiplexed qPCR assay to robustly quantitate and distinguish cell free DNA integrity and concentration in blood plasma and serum is described. The multiplexed system for characterizing cancer in humans includes a sample of serum, plasma, urine, or other biological fluid, the sample comprising cell free DNA, the cell free DNA comprising long and short retrotransposable element targets and an added internal positive control, the long and short targets being independent of each other, a distinctly labeled TaqMan probe corresponding to each target, a forward primer and a reverse primer corresponding to each target, a DNA standard for generating standard curves, a qPCR system for amplifying the targets and a qPCR data analysis system. The assay provides an accurate, minimally-invasive, rapid, high-throughput, and cost-effective method with the potential to complement or replace existing methods for detection, diagnosis, prognosis, treatment monitoring and / or surveillance of cancer, thereby improving patient outcomes.

The application provides an On-DNA-gamma-hydroxylamine compound and a preparation method thereof, and belongs to the field of gene coding compound library construction. The On-DNA-gamma-hydroxylamine compound is a compound shown in formula I. The method for synthesizing the On-DNA-gamma-hydroxylamine compound has small DNA damage, good universality, simple operation and mild conditions. The On-DNA-gamma-hydroxylamine compound has high yield, and the DNA integrity in the product is good. The application enriches the chemical reaction type for synthesizing the coding compound library on DNA, establishes a method for constructing a gene coding compound library with a new gamma-hydroxylamine skeleton, and has a very good application prospect in lead drug development.

The invention discloses a preparation method and application of a magnetic composite adsorption material for efficient enrichment of DNA in a water environment, and belongs to the technical field of environmental monitoring and sample pretreatment. The material takes porous diatomite microspheres as a matrix, ferroferric oxide nanoparticles are loaded in the pores in situ, polyethyleneimine with the weight-average molecular weight of 10000-70000 Da is covalently grafted on the surface, and the material has high adsorption capacity and selectivity. A column tube is filled with the material to form a solid-phase extraction column, and the solid-phase extraction column is matched with an eluent which contains high-concentration NaCl and has the pH value of 8.0-8.5, so that efficient enrichment by an adsorption-elution two-step method can be realized. The method provided by the invention solves the problems of low adsorption capacity, poor recovery rate and tedious operation when the ultralow-concentration DNA is enriched from a large-volume environmental water sample, has the advantages of simple flow, high enrichment multiple, good DNA integrity and the like, and is particularly suitable for environmental DNA monitoring.