Solid-phase substrate as well as treatment method and application thereof

A substrate and relative humidity technology, applied in the field of nucleic acid chips, can solve the problems of difficulty in controlling the stability of nucleic acid molecule distribution density and instability, and achieve the effects of improving sequencing quality, reducing non-specific adsorption, improving distribution density and stability

Active Publication Date: 2020-05-05
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 substrate as well as treatment method and application thereof
  • Solid-phase substrate as well as treatment method and application thereof
  • Solid-phase substrate as well as treatment method and application thereof

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

The invention discloses a solid-phase substrate as well as a treatment method and application thereof. The solid-phase substrate is provided with at least one silanized surface, wherein the surface isa surface exposed in a specific environment for a certain period of time, the specific environment is an inert gas environment, the temperature of the specific environment is selected from 37-60 DEGC, and the certain period of time is no less than 2 hours. By adopting the solid-phase substrate, the distribution density and stability of nucleic acid molecules immobilized on the surface of the solid-phase substrate are improved, the quality of a chip can be improved, and sequencing quality can be improved when the solid-phase substrate is applied to 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 Applications(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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