DNA extraction reagent, kit and extraction method

Abstract

The invention discloses a DNA extraction reagent, a kit and an extraction method. The invention firstly provides the DNA extraction reagent which comprises a lysis solution, a rinsing solution I, a rinsing solution II and an eluent, wherein the lysis solution comprises guanidine isothiocyanate, Tris-HCl, ethylene diamine tetraacetic acid (EDTA), NaCl, sodium dodecyl sulfate (SDS) and a protease inhibitor. The invention further provides the kit containing the DNA extraction reagent and the method for extracting DNA by using the DNA extraction reagent or the kit. The DNA extraction method is rapid, simple and convenient, is favorable for crushing fresh tissues or frozen tissues, is particularly suitable for rapid operation in surgery, has low relative extraction cost, has high quality of extracted nucleic acid, and can meet the detection requirements of qPCR, NGS, SNP typing and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a DNA extraction reagent, kit and extraction method. Background technique [0002] Nucleic acid amplification and detection technology is widely used, especially in the fields of disease diagnosis, microorganism and pathogenic substance detection. In the field of disease diagnosis, with the advent of the era of precision medicine, genetic testing technology is a necessary technical means for genetic disease diagnosis, prenatal testing, molecular typing and personalized medicine. In the diagnostic process, nucleic acid analysis has many advantages over traditional methods, such as enzyme or antibody analysis, with high sensitivity, high flexibility and short time. [0003] In the diagnostic process, nucleic acid extraction and purification is the basis of nucleic acid detection, and the quality of extracted and purified nucleic acid has an important impact on the accuracy...

AI Technical Summary

Problems solved by technology

Among them, the alkali cracking method usually uses corrosive sodium hydroxide to denature and then neutralizes the precipitation with acid; the phenol extraction method has complicated steps, and organic solvents such as phenol and chloroform are used for extraction, which has high chemical toxicity; the magnetic bead method uses magnetic beads Adsorption and solvent elution methods are used to extract DNA. The operation process requires the use of magnetic sleeves and magnets for adsorption, which limits the operating flux. It usually needs to be completed with the help of a fully automatic magnetic bead extractor; silica gel membrane spin column, using polymer resin membrane interception purification DNA, the whole process requires multiple high-speed centrifugation tube changes, which takes a long time

[0004] The main bottleneck in the widespread use of molecular diagnostics in medical laboratories is the requirement to purify nucleic acids from samples: commercial biological nucleic acid DNA release extraction kits generally use silica gel membrane spin columns and magnetic beads, and the operation process is complicated and time-consuming. At least It takes 1.5 hours, which is not conducive to rapid detection, especially in the detection of disease (cancer) treatment surgery. Therefore, it is urgent to develop a simple, fast and high-quality nucleic acid extraction method

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