6 results about "Heat denaturation" patented technology

This invention discloses a method for repairing DNA damage in FFPE samples, comprising: S1. subjecting FFPE sample DNA to heat denaturation; S2. allowing the DNA to fully renature by slow cooling or ice bath; S3. using random primers to bind to large nicked regions of the DNA fragment; S4. adding a mixed repair enzyme to repair the DNA damage; S5. ligating the DNA repaired in step S4 to obtain large DNA fragments, which are then purified with magnetic beads to obtain the repaired DNA. By using random primer single-strand extension to assist the enzyme reaction system, the efficiency of DNA damage / deletion repair is improved; by using random ligation to assemble fragmented DNA into large DNA fragments, the problem of ineffective amplification caused by excessive DNA fragmentation is solved, without affecting the microarray detection results. The entire DNA repair reaction is performed continuously, without the need for multiple purification operations. Compared with other DNA repair methods, it is simple to operate and low in cost, significantly improving the quality of FFPE sample DNA and increasing its detection pass rate in Infinium CytoSNP-850K whole genome microarray detection.

The application discloses a probiotic composition and a production process of a probiotic fermented bread crumb, and belongs to the technical field of bread crumb processing. The probiotic composition is a microcapsule structure containing malt dextrin, arabic gum and heat-denatured whey protein isolate as wall materials, and lactobacillus acidophilus and lactobacillus rhamnosus as core materials. The structure can ensure that a small amount of live bacteria and metabolic products are slowly released through the micropores of the wall materials in a normal temperature fermentation environment, so that the free lactic acid bacteria are prevented from rapidly proliferating, excessively producing acid and competing with yeast for carbon and nitrogen sources in the initial stage of fermentation. The probiotic composition can provide nutrition for the yeast in the fermentation process, promote cell division and metabolic activity of the yeast, and the polydextrose, inulin and arabic gum in the microcapsule wall materials are hydrophilic colloids, which can form a uniform hydration network in the dough.

This invention relates to the field of biodetection technology, specifically to a non-isolated digital multiplex detection method based on a DNA walker and TdT enzyme amplification. The method first utilizes coding microspheres with capture elements coupled to their surface to specifically bind to target miRNAs or proteins in the sample; then, specific guide DNA (gDNA) corresponding to the target molecule is released through enzymatic digestion or thermal denaturation; the gDNA then... Pf Under the action of Ago endonuclease, the DNA walker on the surface of the encoded microspheres is activated, cyclically cutting the orbital strand to generate multiple DNA fragments with 3'-hydroxyl ends. Finally, TdT enzyme is used to extend fluorescently labeled nucleotides at the hydroxyl ends, achieving in-situ signal amplification, and digital quantitative analysis is performed on the fluorescence signal of the encoded microspheres. This invention achieves high-sensitivity detection of multiple targets in a single reaction without chamber isolation, with detection limits for each target molecule reaching the fM or fg / mL level, providing an effective technical means for the early screening of major diseases such as Alzheimer's.

The present application relates to CuZrO2 NPs hydrolysis nano-enzyme and its preparation method and application, first adopt the coprecipitation method synthesis CuZr (OH) 2 NPs, high temperature calcination, make the intramolecular dehydration synthesis CuZrO2 NPs.The application of CuZrO2 NPs hydrolysis nano-enzyme in proteolysis, mix CuZrO2 NPs with ultrapure water, make CuZrO2 NPs disperse uniformly, then add protein solution, water bath reaction, realize the proteolysis.CuZrO2 NPs still has hydrolysis effect for heat denatured and surfactant denatured protein, and has good recovery rate and wide pH activity range, the selected CuZrO2 NPs is applied to the hydrolysis of insoluble soybean protein and periplaneta americana protein.CuZrO2 NPs has great potential in biomedical and proteomics applications, further widening the possibility of developing nano-protease based on bimetallic oxide nanomaterials.

The application provides a high-efficiency extraction method of animal skin enzymatic gelatin. The method is characterized in that the expansion degree and thermal denaturation law of collagen fibers of animal skin are controlled through the synergistic effect of acid, alkali, salt and heat, the exposure degree of the specific site of protease in the collagen fibers is controlled, the endo-type hydrolysis of the protease on the collagen fibers is promoted, the gelatin extraction rate and gel strength are improved, and the performance of the gelatin can meet the quality standard of industrial gelatin without desalting treatment. The method can realize one-time extraction of animal skin gelatin, the gelatin yield is more than 90%, the gel strength is more than 200 Bloom g, the production efficiency of the gelatin can be obviously improved, and good energy-saving and emission-reducing effects are achieved. The method has important significance for developing the high-efficiency preparation technology of high-performance animal skin gelatin.

This application discloses a method for preparing macromolecular collagen from poultry eggshell membranes. The method includes: pulverizing the poultry eggshell membrane, mixing it with a reducing agent and urea, and intermittently sonicating it under gentle stirring to dissolve and remove keratin; extracting the precipitate with citric acid and pepsin, adjusting the pH of the supernatant to neutral, adding ammonium sulfate for precipitation, dialysis after dissolving the precipitate, and freeze-drying to obtain macromolecular collagen from the poultry eggshell membrane. This application selectively removes keratin under non-thermal, neutral conditions, preserving the natural conformation of the collagen. The obtained collagen exhibits a strong absorption peak at 235 nm and a shoulder peak at 280 nm in its ultraviolet spectrum, with a thermal denaturation temperature of 75 °C, and possesses a complete triple helix structure. This method is mild and yields high results, filling a technological gap in the extraction of macromolecular collagen from poultry eggshell membranes. The obtained product can be used in fields such as biological scaffold materials, tissue engineering, and drug carriers.