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Solid-phase metal ion chelating agent as well as preparation method and application thereof

A metal ion, chelating agent technology, applied in DNA preparation, recombinant DNA technology and other directions, can solve the problems of low content of microorganisms, great influence on activity, increase the difficulty of nucleic acid, etc., and achieve the effect of improving activity

Active Publication Date: 2021-06-29
山西奥睿基赛生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, soil samples are diverse and complex, which leads to the extraction of soil DNA has always been a difficult point in the field of nucleic acid extraction
In nutrient-rich soil, the difficulty of nucleic acid extraction is often increased due to the influence of humic acid; in barren soil, the content of microorganisms is low, and some soil nucleic acids increase the difficulty of nucleic acid extraction to a certain extent.
In addition, in soil samples in extreme environments, such as soil samples in mining areas, there are various metal ions that have a great influence on the activity of enzymes in the process of cracking microorganisms. After adding EDTA in the cracking process, various metal ions can be chelated, but at the same time The activities of most enzymes (such as cellulase and protease in the commonly used compound enzyme helicase) also have adverse effects

Method used

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  • Solid-phase metal ion chelating agent as well as preparation method and application thereof
  • Solid-phase metal ion chelating agent as well as preparation method and application thereof

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preparation example Construction

[0017] The embodiment of the present invention provides a kind of preparation method of solid-phase metal ion chelating agent, specifically comprises the following steps:

[0018] S1, esterification grafting reaction

[0019] Through the esterification grafting reaction of ethylenediaminetetraacetic anhydride and dextran, the prepared product can well remove metal ions in soil samples, and has a good market application prospect.

[0020] In the actual operation process, after mixing the organic solvent, dextran and ethylenediaminetetraacetic anhydride, the esterification grafting reaction is carried out under the condition of 55-85°C; the reaction time is 12-48h. The temperature of the mixed solution formed by the organic solvent, the dextran and the ethylenediaminetetraacetic anhydride is raised to 55-85 DEG C, and the esterification and grafting reaction is carried out by keeping it warm.

[0021] In a preferred embodiment, the reaction temperature of the esterification gra...

Embodiment 1

[0035] This embodiment provides a preparation method of a solid-phase metal ion chelating agent, which includes the following steps: adding 0.5 g of G-25, 0.75 g of ethylenediaminetetraacetic anhydride, and N, N dimethylformamide into a 50 mL two-necked bottle 13.5mL, add a magnet, insert a thermometer, install and fix the reflux tube, heat in a magnetic stirring constant temperature bath, set the temperature at 85°C, and react for 12 hours to terminate the reaction. Take out reactant, suction filter and use 15mL 7% sodium bicarbonate solution to process reaction product, carry out hydrolysis treatment 40 minutes to residual acid anhydride group, follow-up continuation ethanol washing process (when carrying out titration method to measure grafting effect, should be in ethanol Acidify before washing). After washing with absolute ethanol for 5 times, drying at 65°C to evaporate residual ethanol, the grafting efficiency was measured. After measuring the grafting efficiency, the ...

Embodiment 2

[0038]This embodiment provides a preparation method of a solid-phase metal ion chelating agent, which includes the following steps: adding 4.6 g of G-75 and 10.2 g of ethylenediaminetetraacetic anhydride into a 250 mL two-necked bottle, adding N, N dimethylethylene Add 187 mL of amide, add a magnet, insert a thermometer, install and fix the reflux tube, heat in a magnetic stirring constant temperature bath, set the temperature at 55°C, and react for 30 hours to terminate the reaction. Then take out the reactant, suction filter and use 15mL 7% sodium bicarbonate solution to process the reaction product, carry out the hydrolysis treatment 40 minutes to the residual anhydride group, follow-up and continue the ethanol washing process (when carrying out the titration method to measure the grafting effect, should be in acidified before washing with ethanol). Wash with absolute ethanol for 5 times, dry and volatilize the residual ethanol at 65°C, and measure the grafting efficiency. ...

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Abstract

The invention discloses a solid-phase metal ion chelating agent as well as a preparation method and application thereof, and relates to the technical field of chelating agents. The solid-phase metal ion chelating agent is obtained by carrying out esterification grafting reaction on ethylenediamine tetraacetic anhydride and glucan; and preferably, the grafting rate is 20 to 30 percent based on the percentage of the carboxyl group. The prepared product can effectively remove metal ions in a biological sample, can effectively improve the activity of nucleic acid in a soil sample, especially an extreme soil sample, and can be applied to soil DNA extraction.

Description

technical field [0001] The invention relates to the technical field of chelating agents, and in particular to a solid-phase metal ion chelating agent, its preparation method and application. Background technique [0002] Soil microorganisms are one of the important components of the soil ecosystem, and almost all soil processes are directly or indirectly related to soil microorganisms. At present, soil microbial metagenomics is a powerful method for soil microbial research. This method has high sensitivity and accuracy. The premise of using this method is that a certain concentration of high-quality soil genomic DNA can be obtained. [0003] However, soil samples are diverse and complex, which leads to the extraction of soil DNA has always been a difficult point in the field of nucleic acid extraction. In nutrient-rich soil, the difficulty of nucleic acid extraction is often increased due to the influence of humic acid; and in barren soil with low microbial content and some...

Claims

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Application Information

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IPC IPC(8): C08B37/02C12N15/10
CPCC08B37/0009C08B37/0024C12N15/1003Y02P10/20
Inventor 韩磊庞尔国
Owner 山西奥睿基赛生物科技有限公司
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