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Single-stranded DNA nucleic acid modified chitosan magnetic microsphere preparation method

A magnetic microsphere, nucleic acid modification technology, applied in chemical instruments and methods, other chemical processes, etc., to achieve good biocompatibility, reduce detection limit, and overcome the effect of easy shedding

Active Publication Date: 2014-08-20
SOUTH CHINA NORMAL UNIVERSITY
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Problems solved by technology

[0007] Compared with the extensive research and application of chitosan magnetic microspheres and nucleic acid aptamers in their respective related fields, combining chitosan magnetic microspheres and nucleic acid aptamers in solid phase microextraction Sugar magnetic microspheres provide carriers with large specific surface area, good magnetic responsiveness and biocompatibility, and use nucleic acid aptamers as recognition functional units to improve selectivity, and are applied to trace alkaloids, antibiotics or nuclei in biological samples The high selectivity, rapid separation and enrichment of glycoside substances have not been reported in the literature at home and abroad

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  • Single-stranded DNA nucleic acid modified chitosan magnetic microsphere preparation method
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  • Single-stranded DNA nucleic acid modified chitosan magnetic microsphere preparation method

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Embodiment Construction

[0026] This embodiment takes adenosine as an example to describe the present invention in detail, but this does not limit the protection scope of the present invention.

[0027] Such as figure 1 As shown, the modified CS / Fe based on adenosine nucleic acid aptamer 3 o 4 The preparation method of magnetic microspheres is as follows:

[0028] (1) Accurately weigh 14.04g FeCl 3 ·6H 2 O, dissolve with boiled and cooled deionized water, transfer to a 500mL three-necked flask, wash the beaker with boiled and cooled deionized water and transfer all to the three-necked flask to obtain a brownish red solution, feed nitrogen for 5min, add 7.19g FeSO 4 ·7H 2 O mechanically stirred to dissolve. Transfer the three-neck flask to a 90°C water bath, add 40mL of 25% ammonia water drop by drop with a separatory funnel, black precipitate forms, and continue stirring at constant temperature for 1h. For magnetic separation, place the product on a magnet. After 5 minutes, the lower layer is a...

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Abstract

The present invention discloses a single-stranded DNA nucleic acid modified chitosan magnetic microsphere preparation method, which comprises: (1) adopting a chemical co-precipitation method to obtain Fe3O4 magnetic nanoparticles; (2) adopting an inverse emulsion cross-linking method to obtain CS / Fe3O4 magnetic microspheres; (3) adding the CS / Fe3O4 magnetic microspheres to APTES to obtain -NH2 modified CS / Fe3O4 magnetic microspheres; (4) adding DTPA to obtain -COOH modified CS / Fe3O4 magnetic microspheres under effects of EDC and NHS; and (5) adding EDC and other solutions, adding a bovine albumin solution, and shaking to obtain the adenosine aptamer modified CS / Fe3O4 magnetic microspheres. According to the present invention, the disadvantages of easy shedding, instability and the like of the conventional physical adsorption method are overcome, and the immobilization amount of the aptamer is significantly increased.

Description

technical field [0001] The invention relates to the field of chemical analysis and testing, in particular to a single-stranded DNA nucleic acid aptamer modified chitosan / Fe 3 o 4 Preparation method of magnetic microspheres. Background technique [0002] The first step of sample analysis is sample pretreatment. Sample pretreatment is the most critical link in the entire sample analysis process. The required time accounts for about 60% of the entire analysis time, and the possible errors account for more than 30% of the entire analysis error. , which brings disadvantages to the analysis work. Traditional sample pretreatment techniques such as Soxhlet extraction, liquid-liquid extraction, column chromatography, etc., generally have problems such as low efficiency, time-consuming, large amount of toxic organic solvents and cumbersome operations, so the development of efficient, fast, simple and green sample pretreatment Technology is especially important. Biological samples ...

Claims

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Application Information

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IPC IPC(8): B01J20/286B01J20/30
Inventor 胡小刚伍莉莉柳晓飞郝丽贤
Owner SOUTH CHINA NORMAL UNIVERSITY
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