Kit for extracting stable bacterial genome DNA by paramagnetic particle method and extraction method thereof

Abstract

The invention provides a kit for extracting stable bacterial genome DNA by a paramagnetic particle method and an extraction method thereof. The kit comprises a lysis reagent group for lysing and releasing bacterial genome DNA and an extraction reagent group for extracting the genome DNA, wherein the lysis reagent group consists of a buffer solution and a lysis solution; the extraction reagent group consists of a paramagnetic particle binding solution, a washing solution and an eluent; and the paramagnetic particle binding solution consists of sodium salt sodium citrate or sodium acetate, a reducing agent dithiothreitol, guanidine hydrochloride and a paramagnetic particle suspension. The raw materials are reasonably compounded to obtain the optimal proportion and the combined extraction conditions of a paramagnetic particle binding solution compounding system, and the extraction method is optimized, so that the extracted bacterial genome DNA has high purity and complete fragments, is difficult to degrade, has stability far superior to that of the commercially available kit, and has remarkable advantages in subsequent experiments such as DNA storage, hybridization, PCR and the like; and meanwhile, the reagents are nontoxic and have small environmental pollution.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a kit for extracting stable bacterial genomic DNA by a magnetic bead method and an extraction method thereof. Background technique [0002] Nucleic acid extraction and purification technology is the key technology in the application of molecular biology technology in clinical testing at present. The routinely used commercial bacterial genome extraction kit is complicated to operate. It takes more than ten steps from collecting bacteria to obtaining genomic DNA, and the extraction process The adsorption efficiency of genomic DNA is low. The magnetic bead DNA purification method is based on the specific adsorption between polynucleotides and negatively charged magnetic beads on the surface under the condition of high concentration of salt ions, so as to achieve the purpose of DNA purification; this method has high extraction efficiency and simple operation , less steps, ...

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Problems solved by technology

[0006] However, none of the above-mentioned kits used biological reagents capable of stabilizing nucleic acids, nor did they discuss the stability of the extracted genomic DNA, and could not provide effective preliminary guarantees for subsequent biological experiments such as amplification and genetic information analysis.

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