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Preparation method of linear hydrogel containing azide functional groups

A functional group, hydrogel technology, applied in the field of preparation of linear hydrogels, to achieve the effect of easy operation and good controllability

Active Publication Date: 2020-05-19
岱纳科技(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no particularly suitable hydrogel for immobilizing nucleic acids for gene sequencing

Method used

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  • Preparation method of linear hydrogel containing azide functional groups

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[0024] The invention provides a method for preparing a linear hydrogel containing N3 functional groups, which has a simple process and is suitable for production. At the same time, the hydrogel has good biocompatibility, and can effectively realize the immobilization of the hydrogel on the modified chip and Nucleic acid molecules are immobilized on hydrogels.

[0025] A method for preparing a linear hydrogel containing N3 functional groups provided by the present invention comprises the following steps:

[0026] (1) Dissolve acrylamide monomer and methacrylamide azide monomer in water, stir to dissolve and mix well, drum Ar for 15min;

[0027] (2) add water-soluble initiator potassium persulfate and TEMED in the system of step (1), continue to drum Ar5min;

[0028] (3) The system in step (2) was reacted at room temperature for 24 hours, and collected by isopropanol precipitation to obtain a linear hydrogel containing N3 functional groups with a molecular weight of 5,000-100,0...

Embodiment 1

[0058] 1) Dissolve 200mg of acrylamide and 50mg of methacrylamide azide monomer in 10ml of water, stir for 5 minutes to dissolve, add to a 50ml centrifuge tube, the temperature is 25°C, drum Ar for 15min;

[0059] 2) Dissolve 30mg of potassium persulfate in 1ml of water, add 100ml of potassium persulfate solution to the above system, keep the temperature at 25°C, and continue to blow Ar for 5min;

[0060] 3) Add 10 ul of TEMED (tetramethylethylenediamine) to the above system, the temperature is 25°C, and stop blowing;

[0061] 4) The above system was sealed and reacted at room temperature (25° C.) for 24 hours to obtain the target linear hydrogel containing N3 functional groups.

[0062] 5) Add the product to 200ml IPA drop by drop with a plastic dropper while stirring the reacted hydrogel, continue stirring for 5 minutes after the dropwise addition, wait for the floc to settle and separate from the upper solution, and slowly suck out the upper white liquid , Pour the remaini...

Embodiment 2

[0064] 1) Dissolve 200mg of acrylamide and 70mg of methacrylamide azide monomer in 15ml of water, stir for 5 minutes to dissolve, add to a 50ml centrifuge tube, the temperature is 25°C, drum Ar for 15min;

[0065] 2) Dissolve 20mg of potassium persulfate in 1ml of water, add 100ml of potassium persulfate solution to the above system, keep the temperature at 25°C, and continue to blow Ar for 5min;

[0066] 3) Add TEMED 7ul to the above system, the temperature is 25°C, and stop blowing;

[0067] 4) The above system was sealed and reacted at room temperature (25° C.) for 24 hours to obtain the target linear hydrogel containing N3 functional groups.

[0068] 5) Add the product to 200ml IPA drop by drop with a plastic dropper while stirring the reacted hydrogel, continue stirring for 5 minutes after the dropwise addition, wait for the floc to settle and separate from the upper solution, and slowly suck out the upper white liquid , Pour the remaining precipitate into a 50ml centrif...

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Abstract

The invention relates to a preparation method of a linear hydrogel containing azide functional groups. The hydrogel containing the azide functional groups can be connected with nucleic acid moleculesthrough click chemical reaction, and has important application in the field of gene sequencing. The preparation method mainly comprises the following steps: mixing an acrylamide monomer and an acrylamide azide monomer; adding a water-soluble initiator, wherein the water-soluble initiator is a redox initiation system composed of potassium persulfate and TEMED; wherein the mass ratio of the initiator to the acrylamide monomer is 0.01 to 0.1; and then carrying out a reaction for 10-30 h at a temperature of 15-30 DEG C, and carrying out isopropanol precipitation and collection so as to obtain thelinear hydrogel containing the azide functional groups.

Description

technical field [0001] The invention relates to a preparation method of a linear hydrogel containing an azide functional group, which can effectively immobilize nucleic acid molecules and belongs to the field of gene sequencing. Background technique [0002] Hydrogel is a kind of gel with hydrophilic three-dimensional network structure. Hydrogels have good hydrophilicity and are suitable for aqueous reaction systems. Hydrogels are widely used in biology, chemistry and other fields. There are many synthetic methods of hydrogels, and the synthetic methods directly affect the performance of hydrogels. Hydrogels have the advantages of good biocompatibility, low immune response, and potential bioactive base sequences in the structure, and have been widely developed for regenerative medicine, drug delivery, and tissue adhesion, and can also be designed for It has been widely used in 3D cell culture for assembling extracellular matrix to provide three-dimensional support for cel...

Claims

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Application Information

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IPC IPC(8): C08F220/56C08F220/58C08J3/075C08L33/26C12Q1/6869
CPCC08J3/075C08F220/56C12Q1/6869C08J2333/26C12Q2565/518C08F220/60
Inventor 刘海涛段海峰陈子天乔朔康力
Owner 岱纳科技(北京)有限公司
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