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Solid phase substrates, their processing methods and uses

A substrate and relative humidity technology, applied in the field of nucleic acid chips, can solve the problems of instability and difficult to control the stability of the distribution density of nucleic acid molecules, and achieve the effects of reducing non-specific adsorption, improving sequencing quality, and improving interference.

Active Publication Date: 2022-03-25
GENEMIND BIOSCIENCES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the distribution density and stability of nucleic acid molecules are still difficult to control and unstable.

Method used

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  • Solid phase substrates, their processing methods and uses
  • Solid phase substrates, their processing methods and uses
  • Solid phase substrates, their processing methods and uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Placed at 57°C after silanization in Example 1

[0090] (1) cleaning the glass substrate: cleaning the surface with 5% (mass concentration) hydrochloric acid for 3 hours;

[0091] (2) Activated glass substrate: soak in piranha solution (piranha solution) for 30 minutes, wash 5 times with ultrapure water, and wash 1 time with ethanol (surface hydroxylation);

[0092] (3) Surface silane derivatization reaction: Prepare 2% (mass concentration) GOPTS (3-(2,3-glycidyloxy)propyltrimethoxysilane) ethanol solution, react at 37°C for 5 hours, and then purify Wash with water, 5 times of ethanol, 1 time of acetone, blow dry with nitrogen, and set aside;

[0093] (4) Low-temperature placement treatment: place the prepared chip in a nitrogen drying cabinet, placement conditions: temperature 57°C, relative humidity 55%, placement time 4 days;

[0094] (5) Immobilization of amino-DNA-CY3 molecules: Prepare 0.25M phosphate buffer solution containing amino-DNA-CY3 molecules, spread on t...

Embodiment 2

[0096] Placed at 47°C after silanization in Example 2

[0097] (1) cleaning the glass substrate: cleaning the surface with 5% (mass concentration) hydrochloric acid for 3 hours;

[0098] (2) Activated glass substrate: soak in piranha solution for 30 minutes, wash with ultrapure water for 5 times, and wash with ethanol once;

[0099] (3) Surface silane derivatization reaction: prepare 2% (mass concentration) GOPTS ethanol solution, react at 37°C for 5 hours, then wash with pure water 5 times, ethanol 5 times, and acetone once, blow dry with nitrogen, and set aside;

[0100] (4) Low-temperature placement treatment: place the prepared chip in a nitrogen drying cabinet, placement conditions: temperature 47°C, relative humidity 55%, placement time 4 days;

[0101] (5) Immobilization of amino-DNA-CY3 molecules: Prepare 0.25M phosphate buffer solution containing amino-DNA-CY3 molecules, spread on the surface, react at 37°C for 10 hours, and then cross-wash with 1XPBS (pH=7.4) and pu...

Embodiment 3

[0103] Example 3 Placed at 37°C after silanization

[0104] (1) cleaning the glass substrate: cleaning the surface with 5% (mass concentration) hydrochloric acid for 3 hours;

[0105] (2) Activated glass substrate: soak in piranha solution for 30 minutes, wash with ultrapure water for 5 times, and wash with ethanol once;

[0106] (3) Surface silane derivatization reaction: prepare 2% (mass concentration) GOPTS ethanol solution, react at 37°C for 5 hours, then wash with pure water 5 times, ethanol 5 times, and acetone once, blow dry with nitrogen, and set aside;

[0107] (4) Low-temperature placement treatment: place the prepared chip in a nitrogen drying cabinet, placement conditions: temperature 37°C, relative humidity 55%, placement time 4 days;

[0108] (5) Immobilization of amino-DNA-CY3 molecules: Prepare 0.25M phosphate buffer solution containing amino-DNA-CY3 molecules, spread on the surface, react at 37°C for 10 hours, and then cross-wash with 1XPBS (pH=7.4) and pure ...

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Abstract

A solid phase substrate, its processing method and application, the solid phase substrate has at least one silanized surface, the surface is a surface exposed to a specific environment for a certain period of time, the specific environment is an inert gas environment, and the temperature of the specific environment is selected from 37°C to 60°C, a certain period of time is not less than 2 hours. The invention improves the distribution density and stability of the nucleic acid molecules fixed on the surface of the solid phase substrate, improves the chip quality, and can improve the sequencing quality when used in sequencing.

Description

technical field [0001] The invention relates to the technical field of nucleic acid chips, in particular to a solid phase substrate, its processing method and application. Background technique [0002] Chip is an important tool in nucleic acid sequencing, widely used in various types of nucleic acid sequencing, especially the third-generation single-molecule sequencing chip is composed of nucleic acid (such as DNA) fragments randomly fixed on glass and other substrates to form millions of single Molecular points are obtained. The density and stability of nucleic acid molecules are mainly determined by the density and distribution state of the active group coating on the surface of the substrate. [0003] In the prior art, silane molecules are usually used to perform a silanization reaction on the surface of the chip substrate to form a silanized modified surface, that is, an active group coating. The chip substrate after the silylation reaction is generally placed in an in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/34
CPCC12Q1/6874C03C17/30C03C2217/70C03C2218/11C03C2218/31
Inventor 赵智陆永艺王琦颜钦赵陆洋
Owner GENEMIND BIOSCIENCES CO LTD
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