Solid phase substrate, processing method thereof, and method for determining processing conditions

A basic and specific technology, applied in the direction of biomass post-processing, biomass pre-treatment, biochemical equipment and methods, etc., can solve the problems of unconfirmed or failed, inaccurate and misleading accelerated experiments

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

This principle is reliable in most cases, but for special materials such as nucleic acid chips, the use of the above principles will be very misleading or inaccurate, which will lead to uncertainty or failure of accelerated experiments

Method used

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  • Solid phase substrate, processing method thereof, and method for determining processing conditions
  • Solid phase substrate, processing method thereof, and method for determining processing conditions
  • Solid phase substrate, processing method thereof, and method for determining processing conditions

Examples

Experimental program
Comparison scheme
Effect test

experiment example 125

[0260] Experimental example Store the chip at 125°C

[0261] Take a number of chips (more than 12, such as 15) from the same batch and perform the following processing:

[0262] (1) cleaning the glass substrate: cleaning the surface with 5% hydrochloric acid for 3 hours;

[0263] (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;

[0264] (3) Surface silane derivatization reaction: prepare 2% GOPTS (3-(2,3-epoxypropoxy)propyltrimethoxysilane) ethanol solution, react at 37°C for 5 hours, then 5 times with pure water, 5 times Wash with ethanol and acetone once, blow dry with nitrogen, and set aside;

[0265] (4) Low-temperature storage treatment: store the prepared chips in a nitrogen cabinet at a temperature of 25°C and a relative humidity of 55%, and store them for different periods of time (0 days, 60 days, 180 days, 360 days, 720 days);

[0266] (5) Immobilization of a...

experiment example 257

[0269] Experimental example Store the chip at 257°C

[0270] Take a number of chips (more than 12) from the same batch and perform the following processing:

[0271] (1) cleaning the glass substrate: cleaning the surface with 5% hydrochloric acid for 3 hours;

[0272] (2) Activated glass substrate: soak in piranha solution for 30 minutes, wash with ultrapure water for 5 times, and wash with ethanol once;

[0273] (3) Surface silane derivatization reaction: prepare 2% GOPTS ethanol solution, react at 37°C for 5 hours, then wash 5 times with pure water, 5 times with ethanol, 1 time with acetone, blow dry with nitrogen, and set aside;

[0274] (4) Low-temperature storage treatment: store the prepared chips in a nitrogen cabinet at a temperature of 57°C and a relative humidity of 55%, and store them for different periods of time (0 days, 2 days, 4 days, 7 days);

[0275] (5) Immobilization of amino-DNA-CY3 molecules: Take out several chips (for example, 3) on day 0, 2, 4, and 7 ...

Embodiment 3

[0282] Preservation at 25°C after silanization in Example 3

[0283] (1) cleaning the glass substrate: cleaning the surface with 5% hydrochloric acid for 3 hours;

[0284] (2) Activated glass substrate: soak in piranha solution for 30 minutes, wash with ultrapure water for 5 times, and wash with ethanol once;

[0285] (3) Surface silane derivatization reaction: prepare 2% GOPTS ethanol solution, react at 37°C for 5 hours, then wash 5 times with pure water, 5 times with ethanol, 1 time with acetone, blow dry with nitrogen, and set aside;

[0286] (4) Fluorescence detection: Detect the number of fluorescent points in different areas on the surface of the chip on a Tirf microscope, and the detection wavelength is 530 nm, and the density constant C of the chip when the storage time is 0 is obtained. 0 ;

[0287] (5) Low-temperature storage treatment: store the prepared chips in a nitrogen cabinet, storage conditions: temperature 25°C, relative humidity 55%, storage time 4 days; ...

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Abstract

The invention relates to a solid phase substrate, a processing method thereof, and a method for determining processing conditions. The solid phase substrate is provided with at least one silanized surface; the surface is exposed in specific environment for a certain time and can realize that the amount of nucleic acid molecules fixed on the surface achieves an expected value; the specific environment is inert gas environment, and the temperature of the specific environment is selected from 37 DEG C to 60 DEG C; and the certain time meets a preset relation, and the preset relation is the relation between the duration that the surface is exposed in the specific environment and the amount of the nucleic acid molecules fixed by the surface. Thus, the accuracy of chip aging experiments can be enhanced; and the solid phase substrate can be accurately used for guiding the production or research and development of chips, especially for application scenes with high requirements for the chips.

Description

technical field [0001] The invention relates to the field of chip technology, in particular to a solid phase substrate, a processing method thereof and a method for determining processing conditions. Background technique [0002] Chip is an important tool in nucleic acid detection, widely used in various types of nucleic acid sequencing and nucleic acid detection, 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 hundreds of Tens of thousands of single molecule points were 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. Th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34C12M1/00C40B50/18C40B40/06
CPCC12Q1/6806C12Q1/6874C12Q1/6837C40B50/18C40B40/06
Inventor 赵智高锦鸿颜钦赵陆洋
Owner GENEMIND BIOSCIENCES CO LTD
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