8 results about "Dna recovery" patented technology

The invention discloses a reagent combination for protecting fragment integrity in DNA methylation detection, the reagent combination is used for protecting fragment integrity in DNA methylation detection sample transformation, and the reagent combination comprises tetrahydrofurfuryl alcohol, L-ascorbyl palmitate and ethylenediamine tetraacetic acid; wherein the volume concentration of the tetrahydrofurfuryl alcohol is 10%-80%; the molar concentration of the L-ascorbyl palmitate is 10 mM to 100 mM; the molar concentration of the ethylenediamine tetraacetic acid is 1 mM to 10 mM. The invention also provides a kit and a method for DNA conversion by using the kit. According to the reagent combination, damage of sulfite to DNA in the transformation process can be reduced, DNA degradation is prevented, a transformed fragment is longer than that of a traditional transformation method, the DNA recovery concentration and the library concentration are higher, follow-up detection is facilitated, and the sensitivity of a follow-up detection method is improved.

The utility model relates to a DNA (deoxyribonucleic acid) rapid crude extraction kit based on a paper base, and belongs to the technical field of DNA extraction devices of paper bases. Comprising a cover plate and a bottom groove; the cover plate is detachably connected with the bottom groove; a plurality of groups of circular hollows and a plurality of groups of circular grooves are formed in the cover plate; the circular hollowed-out parts and the circular grooves are opposite in position and consistent in number on the cover plate; a rectangular groove is formed in the bottom groove; a water-absorbing paper base is mounted in the rectangular groove; and a circular DNA recovery paper substrate is arranged in the circular hollow part and is positioned at the upper part of the water-absorbing paper base. According to the kit, the cover plate and the bottom groove are detachably connected, so that the kit is convenient to carry and store; the kit adopts the design of circular hollows and circular grooves, so that the DNA recovery paper substrate is convenient to place and position, complex operation is not needed, and the DNA extraction efficiency is improved; the water-absorbing paper base is adopted to assist liquid transfer, the efficiency is high, DNA does not need to be repeatedly transferred to recover the paper base, and the paper base is effectively prevented from being broken or deformed.

The invention discloses a buffer solution suitable for collecting a biological sample in a low-temperature environment. The buffer solution comprises the following components: an alcohol solvent combination which is selected from at least two of methanol, ethanol and isopropanol; glycerol accounting for 5%-30% of the total mass of the buffer solution; the buffer solution is Tris-HCl, and the concentration of the buffer solution is 5-50 mM; a chelating agent: EDTA with a concentration of 0.5-5 mM; and the balance of double distilled water. The method comprises the following steps: a, mixing and stirring the alcohol solvent combination according to the formula ratio; b, adding glycerol, and uniformly stirring and mixing; c, a buffer solution Tris-HCl, a chelating agent EDTA and double distilled water are sequentially added, and continuous stirring is conducted till complete dissolution is achieved. The buffer solution is used for collecting a metal surface biological sample in an environment of lower than-20 DEG C. The buffer solution provided by the invention can keep liquid flow in an extremely cold environment as low as-80 DEG C, and meanwhile, the DNA recovery rate and the subsequent detection effect of a metal surface biological sample are remarkably improved.

The invention belongs to the field of a trace sample nucleic acid extraction method, and discloses a fresh poultry egg high-efficiency DNA extraction method based on embryonic disc separation, the method adopts an acoustoelectric coupling micropore cracking-separation technology, the method comprises the following steps: synchronously applying 1.5 MHz high-frequency sound wave and a 2 kV / cm, 1 kHz alternating electric field in a micro-fluidic chip, generating controllable nano-scale micropores in an embryonic disc cell membrane, and separating the embryonic disc cell membrane by using an ultrasonic wave; the ultra-fast cracking within 500 microseconds is realized; split products are directionally screened through a polytetrafluoroethylene microporous membrane under the driving of an electric field, only DNA molecules are allowed to pass through, proteins and lipids are intercepted, and thermal damage and chemical residues are avoided; the method is suitable for 0.5 microliter ultramicro embryonic disc samples, and the DNA recovery rate is gt; the purity is 90%, and the purity is A260 / 280gt; 1.8, the requirements of high-throughput sequencing and SNP (Single Nucleotide Polymorphism) analysis are met.

The invention discloses a method for improving biological aerosol sampling efficiency, and relates to a biological aerosol collection technology in the field of environmental science, and the method comprises the following steps: (1) using a quartz glass fiber filter membrane (the size is 250 mm * 230 mm, and the aperture is 2.2 [mu] m) to cooperate with an air sampler to collect microbial particles; (2) physically stripping substances on the surface layer of the hair surface of the filter membrane by adopting an ultraviolet sterilized polyethylene glycol terephthalate (PET) hard brush (the length of brush hair is 2.85-3.15 cm), and collecting microorganisms; and (3) extracting DNA through a soil DNA kit, and combining fluorescence quantification and agarose gel electrophoresis detection. According to the method, a quartz film rough surface-supporting surface layered structure is creatively utilized, efficient collection of microbial particles is achieved through physical stripping, the limitation of a traditional chemical elution method is overcome, and the DNA recovery rate and the operation efficiency are remarkably improved; the sampling efficiency is superior to that of a mixed cellulose ester filter membrane, and the cost benefit and the adaptability of a high-flow sampler are both considered. The method can be widely applied to the fields of environmental monitoring and public health.

The invention belongs to the technical field of biology, and particularly relates to a method for extracting metagenomes of microorganisms in a mud-water mixture. Comprising the following steps: S1, mud-water sample separation: separating mud and a water body part in mud-water; s2, extraction of metagenomes in the soil: extracting the metagenomes in the soil; s3, water body sample separation: separating microorganisms from free DNA in the water body; s4, extracting metagenomes of thallus samples in water, and extracting metagenomes of microorganisms in the water; and S5, extracting metagenome of the free DNA sample in water, and extracting free DNA in the water. According to the method disclosed by the invention, almost all microorganism DNAs can be effectively recovered from the mud-water mixture, and the DNA loss caused by sample separation is reduced. The DNA recovery rate is improved through optimized splitting decomposition and purification steps, and it is ensured that a high-quality DNA sample can be obtained even under extreme conditions.

The invention provides a composition and application of the composition in conversion of DNA (deoxyribonucleic acid) from hydrosulfite, and belongs to the technical field of biology. The oyster glycogen is added into the conversion liquid, so that the loss of DNA in the hydrosulphite conversion and recovery process can be effectively reduced, the recovery efficiency of DNA is improved, and hydrosulphite conversion and methylation detection of low-input DNA are facilitated.

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.