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Preparation method of double cross-linked hydrogel of dna regular tetrahedron-rolling circle amplification product

A rolling circle amplification and regular tetrahedron technology, which is applied in DNA preparation, biochemical equipment and methods, gel preparation, etc., can solve the limitations of the popularization and application of DNA hydrogels, the low mechanical strength of hydrogels, and the difficulty in carrying packages. Buried things, etc.

Active Publication Date: 2021-04-23
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] The method of preparing nucleic acid hydrogel based on RCA technology has the problems of long time-consuming, low hydrogel mechanical strength, and difficult loading of embedding materials, resulting in low availability
At the same time, the microstructure of RCA hydrogel is a single shape of nanoflowers, and it is difficult to control its size and shape during the preparation process, which greatly limits the popularization and application of DNA hydrogels.

Method used

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  • Preparation method of double cross-linked hydrogel of dna regular tetrahedron-rolling circle amplification product
  • Preparation method of double cross-linked hydrogel of dna regular tetrahedron-rolling circle amplification product
  • Preparation method of double cross-linked hydrogel of dna regular tetrahedron-rolling circle amplification product

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Experimental program
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Effect test

Embodiment 1

[0074] Example 1. Preparation and Characterization of DNA Tetrahedron-Rolling Circle Amplification Product Double Crosslinked Hydrogel

[0075] (1) Experimental materials

[0076] The experimental reagent information used in this embodiment is shown in Table 1, and the nucleotide sequences of the designed primers are shown in Table 2 and the sequence list.

[0077]

[0078] Except for the experimental reagents in Table 1, the experimental water was from the Milli-Q pure water system. Other reagents were purchased from Sinopharm Group.

[0079]

[0080] In Table 2, the 5' end of the padlock probe is modified by phosphorylation, and its chemical structure is:

[0081]

[0082] The sequences listed in Table 2 are all artificially synthesized.

[0083] (2) RCA reaction

[0084] 1) Ligation reaction

[0085] Such as figure 1 As shown, the first step of the RCA reaction is to ligate the padlock probes with the help of ligation primers through the action of T4 ligase t...

Embodiment 2

[0111] Example 2, the loading of Dox in the double cross-linked hydrogel of DNA regular tetrahedron-rolling circle amplification product

[0112] (1) Experimental materials

[0113] The experimental reagent information used in this embodiment is shown in Table 6.

[0114] Table 6

[0115] experimental reagent factory level Dox Sigma biological reagent

[0116] Except for the experimental reagents in Table 6, the experimental water was from the Milli-Q pure water system. Other reagents were purchased from Sinopharm Group.

[0117] (2) Loading of Dox in double cross-linked hydrogel of DNA regular tetrahedron-rolling circle amplification product

[0118] 1) Dox loading

[0119] In the present invention, Dox is selected as an example to prove that the DNA regular tetrahedron-rolling circle amplification product double-crosslinked hydrogel has the ability to carry dyes or drugs. Take the absorbance value of Dox at 488 nm as the ordinate, and the con...

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Abstract

The invention establishes a preparation method of double cross-linked hydrogel of DNA regular tetrahedron-rolling circle amplification product. This method combines the fast self-assembled DNA regular tetrahedron (TDN) and rolling circle amplification (RCA) products, and can obtain double cross-linked hydrogels in only 10 minutes, which solves the problem of RCA technology to prepare DNA hydrogels. question of time. At the same time, the double-stranded structure of TDN greatly enhances the mechanical strength of the hydrogel network and provides chimeric sites for the efficient loading of dyes, drugs and other embedded substances. In addition, the double-crosslinked hydrogel microscopically has a honeycomb-like structure formed by dense crosslinking of nanoflowers, and its density and the particle size of nanoflowers can be adjusted by TDN concentration and RCA reaction time. control. The invention realizes the rapid preparation of TDN-RCA product double-crosslinked hydrogel with the advantages of high efficiency of embedding, controllable morphology, strong versatility, etc., and has very good performance in molecular detection, drug loading and delivery, etc. application prospects.

Description

technical field [0001] The invention belongs to the field of biological materials, and in particular relates to a method for preparing a DNA regular tetrahedron-rolling circle amplification product double-crosslinked hydrogel. Background technique [0002] The method of preparing nucleic acid hydrogels based on RCA technology has the problems of long time-consuming, low mechanical strength of hydrogels, and difficulty in carrying embeddings, resulting in low availability. At the same time, the microstructure of RCA hydrogel is a single shape of nanoflowers, and it is difficult to control its size and shape during the preparation process, which greatly limits the popularization and application of DNA hydrogels. The introduction of nucleic acid self-assembled DNA tetrahedron (TDN) advanced structure can effectively make up for the deficiency of RCA technology to prepare nucleic acid hydrogels, and provides a new idea for the preparation of DNA hydrogels. On the one hand, beca...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
CPCA61K9/06A61K31/704A61K47/26A61K49/0052A61P35/00B01J13/0052C12Q1/6844G01N21/31C12Q2531/125C12Q2521/101C12Q2521/501C12Q2525/307
Inventor 许文涛黄昆仑张洋子谢银侠
Owner CHINA AGRI UNIV
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