5 results about "Dna integrity" patented technology

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 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.

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.