Kit for extracting stable bacterial genome DNA by magnetic bead method and its extraction method
An extraction method and genome technology, applied in the field of molecular biology, can solve the problems of undiscussed genomic DNA stability, unused biological reagents, etc., and achieve the effects of promoting aggregation and precipitation, not easy to degrade, and reducing repulsive force.
Active Publication Date: 2022-07-05
WUHAN TEXTILE UNIV
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[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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Embodiment 1
[0045] Embodiment 1 of the present invention provides a kit for extracting stable bacterial genomic DNA by magnetic bead method, which includes a lysis reagent set for lysing and releasing bacterial genomic DNA and an extraction reagent set for extracting genomic DNA; the lysis reagent The group is composed of buffer and lysate; the extraction reagent group is composed of magnetic bead binding solution, washing solution and eluent; the magnetic bead binding solution is composed of sodium salt sodium citrate or sodium acetate, reducing agent dithiothreose alcohol, guanidine hydrochloride and magnetic bead suspension; the washing solution is an isopropanol aqueous solution with a volume fraction of 80%; the eluent is distilled water.
[0046] The buffer solution is a mixed aqueous solution containing 1% triton by mass, 5mM disodium EDTA, 20mM tris hydrochloride and 0.3% lysozyme by mass. The pH of the buffer is The value is 7.5. The lysing solution is a mixed aqueous solution c...
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Abstract
The invention provides a kit for extracting stable bacterial genome DNA by a magnetic bead method and an extraction method thereof. The kit includes a lysis reagent set for lysing and releasing bacterial genomic DNA and an extraction reagent set for extracting genomic DNA; the lysis reagent set is composed of buffer and lysis solution; the extraction reagent set is composed of magnetic bead binding solution, It is composed of washing solution and eluent; the magnetic bead binding solution is composed of sodium salt sodium citrate or sodium acetate, reducing agent dithiothreitol, guanidine hydrochloride and magnetic bead suspension. The present invention obtains the optimal ratio of the magnetic bead binding solution compounding system and the combined extraction conditions by rationally compounding the above-mentioned raw materials, and at the same time optimizes the extraction method, so that the extracted bacterial genomic DNA has high purity, complete fragments, and is not easy to be extracted. Degradation and stability are far better than those of commercial kits, and have significant advantages in subsequent experiments such as DNA storage, hybridization, PCR, etc.; at the same time, the reagents are non-toxic and have little 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 detection. The routinely used commercial bacterial genome extraction kits are complicated to operate. More than ten steps are required from collecting bacteria to obtaining genomic DNA, and the extraction process The adsorption efficiency to genomic DNA is low. The magnetic bead-based DNA purification method is based on the specific adsorption of polynucleotides with 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. , few steps, e...
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IPC IPC(8): C12N15/10
CPCC12N15/1013C12N15/1003
Inventor 钟亚平王栋鲁振坦
Owner WUHAN TEXTILE UNIV