CN107215860B - Preparation method of graphene/DNA ordered self-assembly structure - Google Patents
Preparation method of graphene/DNA ordered self-assembly structure Download PDFInfo
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Abstract
A preparation method of a graphene/DNA ordered self-assembled structure comprises the following steps: (1) soaking and modifying the silicon wafer loaded with the graphene by using a polyglutamic acid or polyacrylic acid solution; (2) and soaking and modifying with a single-stranded DNA solution to form a DNA ordered self-assembled structure on the surface of the graphene. The invention provides a graphene/DNA ordered self-assembly structure with strong universality and functionalization, which is used for replacing a conventional self-assembly structure on a rigid substrate in the prior art, and effectively expands the application of the DNA ordered self-assembly structure in a plurality of fields such as biochemical detection, scientific research and the like.
Description
Technical Field
The invention relates to the field of graphene composite materials, in particular to a graphene/DNA ordered self-assembly structure and a preparation method thereof.
Background
Graphene has a two-dimensional lattice structure, and carbon atoms in a plane are connected by sp2 hybridized orbitals to form a hexagonal lattice structure. Due to its unique physical properties, graphene is considered to have great promise in sensitive electrical detection of chemical or biological molecules.
Before exploring sensitive detectors based on Graphene, a practical modification method capable of functionalizing a Graphene surface is firstly found, and it is reported (Mohanty, n.; Berry, v. Nano L et. 2008, 8, 4469-4476.) that amino-terminated single-stranded DNA is bonded to Graphene oxide through a covalent bond, but the Graphene oxide is not selected, and the ordered assembly of surface DNA is not achieved.
The field needs to develop a method capable of orderly assembling biomolecules on the surface of graphene, so that the integrity of graphene sheets is not damaged, and the biomolecules can be orderly assembled on the surface of graphene.
Disclosure of Invention
The invention aims to provide a graphene/DNA ordered self-assembly structure with strong universality and functionalization by taking graphene as a carrier, so as to replace a conventional self-assembly structure on a rigid substrate in the prior art, and effectively expand the application of the DNA ordered self-assembly structure in various fields such as biochemical detection, scientific research and the like.
Firstly, the invention provides a preparation method of a graphene/DNA ordered self-assembly structure, which is characterized by comprising the following steps:
(1) soaking the silicon wafer loaded with the graphene in a polyglutamic acid or polyacrylic acid solution for a certain time, then respectively cleaning the silicon wafer with deionized water and absolute ethyl alcohol, and then drying the silicon wafer with nitrogen;
(2) soaking the graphene in a single-stranded DNA solution of PBS buffer solution with the solvent of 0.01-0.1M, in which 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride is dissolved, washing the graphene with deionized water and absolute ethyl alcohol respectively, and blow-drying the graphene with nitrogen gas to form a DNA ordered self-assembled structure on the surface of the graphene.
Specifically, the graphene-loaded silicon wafer is stripped on the silicon wafer by a micromechanical force stripping method.
In order to remove surface residues, the vacuum degree of the silicon wafer loaded with the graphene is 1.0 × 10 at 350-600 DEG C-6Pa-1.0×10-4And (4) carrying out vacuum annealing for 2-5h under Pa to remove residual glue on the silicon wafer.
The concentration of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride used was 0.01M to 0.1M. The weight-average molecular weight of the polyglutamic acid or polyacrylic acid solution is 20000-50000, and the concentration is 0.1 mg/ml-2 mg/ml.
Taking efficiency and stability into consideration, and the soaking time in the step (1) is 0.5-2 h. The soaking time in the step (2) is 6-24 h.
Specifically, the single-stranded DNA sequence is AAAAATTTTT. Of course, the DNA sequence involved in the prior art, which is capable of orderly self-assembly, is not limited to the specific DNA, and can be self-assembled on the surface of graphene according to the design concept of the present invention.
Compared with the prior art, the invention has the following technical advantages:
(1) the graphene is creatively introduced to serve as a substrate for DNA self-assembly, and is different from the traditional hard materials such as silicon, silicon dioxide, mica, quartz and the like, and the graphene achieves the effect of a flexible substrate by virtue of a unique single-layer structure;
(2) the graphene is used as a substrate, so that excellent physical properties of the graphene can be effectively combined into a DNA ordered self-assembly structure, the regulation and control of the physical properties of the graphene are realized, particularly the influence of the electron transport performance in the graphene is realized, and the technical effect and the application prospect are beyond imagination.
(3) As a general preparation method, the application is wide in application, and especially the existing functionalized DNA sequence has wide significance and prospect in scientific research and practical application by fixing and ordering the DNA sequence.
Detailed Description
The technical solution of the present invention is further explained by the following examples, but the scope of the present invention is not limited in any way by the examples.
Example 1
1. Preparing graphene on a silicon wafer by using a mechanical stripping method, and then loading the graphene-loaded silicon wafer at 350 ℃ and under the vacuum degree of 1.0 × 10-6And (4) carrying out vacuum annealing for 2h under Pa.
2. Soaking the silicon wafer loaded with graphene in polyglutamic acid with the weight-average molecular weight of 20000 and the concentration of 0.1mg/ml for 2h, washing with deionized water and absolute ethyl alcohol respectively, and blow-drying with nitrogen.
3. The DNA fragments were soaked in a single-stranded DNA solution of 0.01M PBS buffer (0.04 mg/ml) containing 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, then washed with deionized water and absolute ethanol, and then dried with nitrogen. The DNA used in the experiment had the sequence AAAAATTTTT, and the concentration of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride was 0.01M.
Example 2
1. Preparing graphene on a silicon wafer by using a mechanical stripping method, and then enabling the silicon wafer loaded with the graphene to be at 450 ℃ and under the vacuum degree of 1.0 × 10-4And (5) carrying out vacuum annealing for 5h under Pa.
2. Soaking the silicon wafer loaded with graphene in polyglutamic acid with the weight-average molecular weight of 40000 and the concentration of 0.5 mg/ml for 1h, then respectively washing with deionized water and absolute ethyl alcohol, and then blowing and drying with nitrogen.
3. The DNA fragments were soaked in a single-stranded DNA solution of 0.05M PBS buffer (0.08 mg/ml solvent) containing 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride for 6 hours, washed with deionized water and absolute ethanol, and then dried with nitrogen. The DNA used in the experiment had the sequence AAAAATTTTT and the concentration of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride was 0.04M.
Example 3
1. Preparing graphene on a silicon wafer by using a mechanical stripping method, and then loading the graphene-loaded silicon wafer at 500 ℃ and under the vacuum degree of 1.0 × 10-6And (4) carrying out vacuum annealing for 2h under Pa.
2. Soaking the silicon wafer loaded with graphene in polyacrylic acid with the weight-average molecular weight of 30000 and the concentration of 1.5 mg/ml for 0.8h, then respectively cleaning the silicon wafer with deionized water and absolute ethyl alcohol, and then blowing the silicon wafer with nitrogen.
3. The DNA fragments were soaked in a single-stranded DNA solution of 0.04M PBS buffer solution containing 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 0.06mg/ml solvent, then washed with deionized water and absolute ethanol, and then dried with nitrogen. The DNA used in the experiment had the sequence AAAAATTTTT, and the concentration of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride was 0.05M.
Example 4
1. Preparing graphene on a silicon wafer by using a mechanical stripping method, and then loading the graphene-loaded silicon wafer at 600 ℃ and a vacuum degree of 1.0 × 10-5And (4) carrying out vacuum annealing for 3h under Pa.
2. Soaking the silicon wafer loaded with the graphene in polyacrylic acid with the weight-average molecular weight of 40000 and the concentration of 1.2 mg/ml for 1.5h, then respectively cleaning the silicon wafer with deionized water and absolute ethyl alcohol, and then blowing the silicon wafer with nitrogen.
3. The DNA fragments were soaked in a single-stranded DNA solution of 0.1M PBS buffer in a 0.1mg/ml solvent containing 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, washed with deionized water and absolute ethanol, and then dried with nitrogen. The DNA used in the experiment had the sequence AAAAATTTTT, and the concentration of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride was 0.1M.
Example 5
1. Preparing graphene on a silicon wafer by using a mechanical stripping method, and then loading the graphene-loaded silicon wafer at 550 ℃ and under the vacuum degree of 1.0 × 10-5And (4) carrying out vacuum annealing for 4h under Pa.
2. Soaking the silicon wafer loaded with graphene in polyacrylic acid with the weight-average molecular weight of 30000 and the concentration of 1.5 mg/ml for 2h, then respectively washing the silicon wafer with deionized water and absolute ethyl alcohol, and then drying the silicon wafer with nitrogen.
3. The DNA fragments were soaked in a single-stranded DNA solution of 0.1M PBS buffer in a 0.2mg/ml solvent containing 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, washed with deionized water and absolute ethanol, and then dried with nitrogen. The DNA used in the experiment had the sequence AAAAATTTTT, and the concentration of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride was 0.05M.
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
Claims (9)
1. A preparation method of a graphene/DNA ordered self-assembly structure is characterized by comprising the following steps:
(1) soaking the silicon wafer loaded with the graphene in a polyglutamic acid or polyacrylic acid solution for a certain time, then respectively cleaning the silicon wafer with deionized water and absolute ethyl alcohol, and then drying the silicon wafer with nitrogen;
(2) soaking the graphene in a single-stranded DNA solution of PBS buffer solution with the solvent of 0.01-0.1M, in which 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride is dissolved, washing the graphene with deionized water and absolute ethyl alcohol respectively, and blow-drying the graphene with nitrogen gas to form a DNA ordered self-assembled structure on the surface of the graphene.
2. The method for preparing the graphene/DNA ordered self-assembled structure according to claim 1, wherein the graphene-loaded silicon wafer is stripped onto the silicon wafer by a micromechanical force stripping method.
3. The method for preparing the graphene/DNA ordered self-assembled structure according to claim 2, wherein the degree of vacuum of the graphene-loaded silicon wafer at 350-600 ℃ is 1.0 × 10-6Pa-1.0×10-4And (4) carrying out vacuum annealing for 2-5h under Pa to remove residual glue on the silicon wafer.
4. The method for preparing the graphene/DNA ordered self-assembled structure according to claim 1, wherein the concentration of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride is 0.01M to 0.1M.
5. The method for preparing the ordered self-assembled graphene/DNA structure as claimed in claim 1, wherein the weight average molecular weight of the polyglutamic acid or polyacrylic acid solution is 20000-50000, and the concentration is 0.1 mg/ml-2 mg/ml.
6. The method for preparing the graphene/DNA ordered self-assembled structure according to claim 1, wherein the soaking time in the step (1) is 0.5h-2 h.
7. The method for preparing the graphene/DNA ordered self-assembled structure according to claim 1, wherein the single-stranded DNA sequence is AAAAATTTTT.
8. The method for preparing the graphene/DNA ordered self-assembled structure according to claim 1, wherein the soaking time in the step (2) is 6-24 h.
9. A graphene/DNA ordered self-assembled structure prepared according to the method defined in any one of claims 1 to 8.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101559944A (en) * | 2009-05-27 | 2009-10-21 | 天津大学 | Conductive graphene film and self-assembly preparation method thereof |
| CN102659918A (en) * | 2012-05-16 | 2012-09-12 | 北京化工大学 | Graphene/dipeptide self-assembly composite film and preparation method thereof |
| CN102875805A (en) * | 2012-09-28 | 2013-01-16 | 浙江理工大学 | Three-dimensional graphene-poly dopamine-gold nanoparticle composite material and preparation method thereof |
| CN102983291A (en) * | 2012-11-27 | 2013-03-20 | 北京大学 | Method for preparing molecular device with surficial biomolecules removed and based on graphene electrodes |
| CN103913496A (en) * | 2014-02-28 | 2014-07-09 | 汕头大学 | Preparation method and application of modified glassy carbon electrode |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101559944A (en) * | 2009-05-27 | 2009-10-21 | 天津大学 | Conductive graphene film and self-assembly preparation method thereof |
| CN102659918A (en) * | 2012-05-16 | 2012-09-12 | 北京化工大学 | Graphene/dipeptide self-assembly composite film and preparation method thereof |
| CN102875805A (en) * | 2012-09-28 | 2013-01-16 | 浙江理工大学 | Three-dimensional graphene-poly dopamine-gold nanoparticle composite material and preparation method thereof |
| CN102983291A (en) * | 2012-11-27 | 2013-03-20 | 北京大学 | Method for preparing molecular device with surficial biomolecules removed and based on graphene electrodes |
| CN103913496A (en) * | 2014-02-28 | 2014-07-09 | 汕头大学 | Preparation method and application of modified glassy carbon electrode |
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