17 results about "Dna integrity" patented technology

The invention discloses a method for analyzing orange peel microbial community structure difference based on a high-throughput sequencing technology, which comprises the following steps: S1, standardized preparation of a sample: collecting orange peel, opening the orange peel, drying in the shade, turning the orange peel and sun-drying until the water content is less than or equal to 12% to obtain an orange peel sample; s2, crushing the orange peel sample, adding a buffer solution containing an impurity adsorbent, and performing water bath and centrifugation to remove flavonoid and polysaccharide impurities; extracting DNA by adopting a DNA extraction kit, and verifying the integrity and purity of the DNA; s3, designing primers for the specific high-resolution sequencing areas of bacteria and fungi, constructing a PCR system by using high-fidelity enzyme, performing amplification, and constructing a sequencing library after recovering target fragments; and S4, obtaining sequence data through a high-throughput sequencing platform, generating a flora feature table, calculating an Alpha diversity index to evaluate the community richness, analyzing and evaluating the community differentiation degree through Beta diversity, and screening core differential flora by adopting a standard that LDAscore is greater than 2. The efficient analysis on the structure difference of the orange peel microbial community is realized.

The invention discloses a method for extracting high-quality genome DNA of peanut leaves, and belongs to the technical field of biological genetic information extraction. The method for extracting the high-quality genome DNA of the peanut leaves comprises the following steps: quickly freezing the peanut leaves, and then grinding; a pretreatment solution is added for pretreatment; centrifuging, discarding the supernatant, adding a CTAB (Cetyltrimethyl Ammonium Bromide) extracting solution, and incubating; after the incubation is finished, extracting; taking supernate, and then carrying out precipitation treatment; centrifuging, and retaining DNA precipitate; and then rinsing and naturally air-drying to obtain high-quality genome DNA (Deoxyribose Nucleic Acid). According to the method, a traditional CTAB method is split, recombined, improved and optimized, the improved CTAB extraction method suitable for the peanuts is established, DNA browning is effectively prevented, high-purity DNA is obtained, meanwhile, DNA integrity, yield and stability are greatly improved, no special requirement for the state of peanut leaves exists, and molecular biology development of the peanuts is greatly promoted.

The invention discloses a depigmentation method suitable for melanoma gene detection, and belongs to the technical field of molecular pathology, the depigmentation method comprises the following steps: pre-treating a melanoma sample, adding into a first part of treating fluid, soaking at 80 DEG C for 20 minutes, adding into a second part of treating fluid, soaking at 80 DEG C for 10 minutes, washing with water, and airing; a preparation method of the first part of treating fluid comprises the following steps: diluting H2O2 with a Tris-HCl buffer solution until the volume concentration of H2O2 is 1%, and then adding NaCl until the mass concentration of NaCl is 1% to obtain the first part of treating fluid; according to the depigmentation method, rapid and efficient depigmentation can be achieved for melanoma with different melanin contents, the DNA integrity and amplification efficiency of the melanoma treated through the depigmentation method are high, and the problem of depigmentation is not prone to occurring.

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.

Disclosed is a composition containing: a heat-treated bacterium having a DIN (DNA Integrity Number) indicating a degree of degradation of 1.0-8.5 of the genomic DNA of the bacterium; a heat-treated bacterium having a Cp value (Crosspoint value) of 15.5-24.0 of the bacterium as measured under prescribed conditions; or a prescribed copy or more of a DNA fragment amplified by a prescribed primer set.

The application discloses a depigmentation method suitable for melanoma gene detection and belongs to the technical field of molecular pathology. The depigmentation method comprises the following steps: after pretreatment of a melanoma sample, the first treatment solution is added, and the sample is soaked at 80 DEG C for 20 min; the second treatment solution is added, and the sample is soaked at 80 DEG C for 10 min; the sample is washed with water and dried; the preparation method of the first treatment solution comprises the following steps: H2O2 is diluted to 1% of the volume concentration of H2O2 by using Tris-HCl buffer solution, then NaCl is added to 1% of the mass concentration of NaCl, and the first treatment solution is obtained; the depigmentation method can realize rapid and efficient depigmentation for melanomas with different melanin contents, the DNA integrity and amplification efficiency of the melanoma treated by using the depigmentation method are high, and the problem of sample loss is not prone to occurring.

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.

The invention provides a detection method based on a DNA integrity index and application, and relates to the technical field of biological detection.In a closed light-operated CRISPR-Cas12a system, a detection sample, terminal deoxyribonucleotide transferase and deoxyribonucleotide triphosphate are subjected to a mixed cycle reaction in advance, a pre-reaction unit is obtained, and on the premise of interference resistance, the DNA integrity index is detected through the pre-reaction unit; the terminal deoxynucleotide transferase specifically recognizes the 3 '-hydroxyl (3'-OH) terminal of the cfDNA and extends a deoxynucleotide chain; after a deoxynucleotide base extends to form a polynucleotide chain sequence, activating a light-operated CRISPR-Cas12a system through ultraviolet rays so as to release crRNA; at the moment, the crRNA recognizes a polynucleotide sequence formed by extension and triggers the trans-cleavage activity of Cas12a; the activated Cas12a further performs non-specific cutting on a single-stranded DNA (ssDNA) fluorescent probe to generate a fluorescent signal, and then the DNA integrity index in the detection sample is calculated according to the fluorescent signal; operation is simple and an anti-interference capability is strong.

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.

The invention discloses a sperm nucleus DNA integrity detection kit, and belongs to the technical field of kits.The sperm nucleus DNA integrity detection kit comprises a reagent detection kit, a cover plate is arranged at the center of the upper end face of the reagent detection kit, protection structures are arranged at the centers of the reagent detection kit and the two side walls of the cover plate, and the four protection structures comprise four protection transverse frames; the four protective cross frames are arranged at the centers of the two side walls of the cover plate and the lower portions of the centers of the two side walls of the reagent detection box respectively, first insertion openings are formed in the front and rear portions of the lower ends of the two side walls of the reagent detection box and the front and rear portions of the centers of the two side walls of the cover plate, and first insertion blocks are arranged at the front and rear portions of the centers of the side walls of the four protective cross frames; besides, when the kit is subjected to external collision or extrusion, external force can be effectively dispersed and buffered, the risk that internal parts such as reagent bottles are broken and damaged due to impact is reduced, the stability of products in the transportation and storage process is enhanced, and operation is simpler, more convenient and more reliable.

The utility model discloses a centrifugal tube upside -down mixed device, related to experimental equipment technical field, including mixing mechanism, holding mechanism and fixed establishment, the holding mechanism at least one side of front and back is provided with for the tensioning mechanism of tensioning fixed establishment, the holding mechanism with the fixed establishment is provided with the guide mechanism for limiting its horizontal direction relative displacement on, beneficial effect lies in: its 60 hole centrifugal tube frame can be compatible with 2ml, 1.5ml centrifugal tube, cooperate automation drive, replace manual batch mixed, promote efficiency and consistency, reduce manpower cost, mechanical buckle and the tensile fastening of reinforcing rib, guide column guide hole restricts horizontal displacement, strengthens stability, avoids sample leakage etc. risk, rubber layer adapts to different tube height, and gentle upside -down mode protects DNA integrity, and conical guide hole and round head guide column make convenient operation, and are applicable to DNA extraction scene etc.

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.