Bacterium nucleic acid extraction solution obtained by magnetic bead method

A technology of bacterial nucleic acid and extract solution, which is applied in the field of molecular biology, can solve the problems of rare earth salt limitations, achieve the effects of reducing dosage, avoiding effect decline, and mild and stable properties

Pending Publication Date: 2020-05-15
重庆市大足区人民医院
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Problems solved by technology

But so far, the application of rare earth salts in biomedicine is still limited to bacteriostats, bactericides and disinfectants, etc.

Method used

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  • Bacterium nucleic acid extraction solution obtained by magnetic bead method
  • Bacterium nucleic acid extraction solution obtained by magnetic bead method
  • Bacterium nucleic acid extraction solution obtained by magnetic bead method

Examples

Experimental program
Comparison scheme
Effect test

preparation example 1-17

[0037] The preparation of preparation example 1-17 bacterial nucleic acid extraction solution

[0038] Described bacterium is preferably Staphylococcus aureus (Latin name Staphyloccocusaureus, bacterial strain number ATCC25923)

[0039] Preparation example 1-17 is the screening of the embodiment of bacterial nucleic acid extraction solution, through the cooperation of different components and conditions, investigate the impact of rare earth elements and surfactants on the extraction of Staphylococcus aureus nucleic acid, the nucleic acid extraction solution components are as shown in the table 1.

[0040] The general preparation method of Preparation Example 1-17 is: 1) a certain amount of rare earth compound (NdCl 3 ), acidic buffer (Tris-HCl at pH 6), surfactant, and chelating agent (EDTA).

[0041] Table 1 Nucleic acid extraction solution components

[0042]

[0043]

[0044] Extract the resuspension of the bacillus cultured overnight by centrifugation and sediment...

Embodiment 1

[0055] The extraction effect of the combined use of rare earth salts and surfactants was explored, and the results are shown in Table 2.

[0056] In order to verify the difference between the extract solution of the present invention and the extract solution combined with various surfactants, one or even several surfactants are replaced with different concentrations of rare earth salts

[0057] In order to compare with a variety of surfactants and extracts, the rare earth salt NdCl 3 Consistent with the concentration of sodium N-lauroyl sarcosinate

[0058] Table 2 Extraction effect

[0059] Preparation example Nucleic acid extraction concentration (ng / uL) OD260 / 280 OD260 / 230 1 32.055 1.55 1.74 2 88.352 1.87 2.03 3 32.428 1.75 1.86 4 38.759 1.78 1.92

[0060] From the extraction of nucleic acid in Preparation Examples 1-4, it can be concluded that only rare earth compounds have no surfactant, and the extraction concentration is ...

Embodiment 2

[0064] On the basis of Preparation Examples 5-7, it was explored that the concentration of nucleic acid extraction was between 35-40ng / uL, and the content of Triton X-100 needed to be added to each of the three extraction solutions. The results are shown in Table 3.

[0065] Table 3 Surfactant dosage

[0066] Preparation example Triton X-100% 5 0.35 6 1.2 7 1

[0067] The results showed that,

[0068] The superposition of three surfactants achieves the same extraction effect. The dosage of Triton X-100 is less than the dosage of Triton X-100 in the superposition of two surfactants, and the dosage of Triton X-100 is the least when used in conjunction with rare earth compounds;

[0069] The addition of rare earth salts can achieve the same extraction effect with less surfactants, that is, the combined use of rare earth salts and surfactants is more efficient than the combined use of surfactants

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Abstract

The invention discloses a bacterium nucleic acid extraction solution obtained by a magnetic bead method. The bacterium nucleic acid extraction solution comprises the following components of a rare earth compound, an acid buffer solution, a surfactant and a chelating agent. Through cooperative usage of the rare earth compound and the surfactant, the bacterium nucleic acid extraction efficiency is improved, and extraction time is shorter.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a bacterial nucleic acid extraction solution by magnetic bead method Background technique [0002] The most classic method in nucleic acid extraction is the phenol-chloroform method, which uses the different solubility of nucleic acid, protein and other substances in different solvents to separate nucleic acid. The phenol-chloroform method is classic and cheap, but it is toxic and takes a long time, which cannot meet the needs of rapid and efficient nucleic acid extraction. In addition, most bacterial extraction reagents on the market use the boiling method. Although the operation is convenient, the yield and purity of nucleic acid are low, which will directly affect the downstream detection results. Magnetic bead nucleic acid extraction is currently a widely used method for rapid nucleic acid extraction. Magnetic particles can bind to specific antibodies, which can specifically...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 董剑席家庄赵峻英潘莉娟谢铌奇徐启峰王杉
Owner 重庆市大足区人民医院
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