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Method for fixing DNA basic group to surface of glass slide

A surface immobilization and glass slide technology, which is applied in the field of immobilizing DNA bases on the surface of glass slides, can solve the problems of expensive reagent preparation, instability, and increased base immobilization time, and achieve high fixation rate, low cost, and wide application. The effect of practicality

Active Publication Date: 2014-11-12
CHINA UNIV OF MINING & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods involve constraints such as high temperature (epoxy surface), expensive preparation reagents (isothiocyanate surface), and instability (mercapto surface).
In addition, in the detection of DNA base fixation rate, fluorescence imaging analysis method is basically used, which requires fluorescent labeling of DNA bases, which greatly increases the time for base fixation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Cut the glass slide into small pieces of 10×10mm, soak in detergent for 5 minutes, rinse with tap water 1-3 times, then ultrasonically clean with deionized water for 5-10 minutes, and rinse with deionized water for 1-3 times; 30%H 2 o 2 , 98%H 2 SO 4 Solution, according to the volume ratio of 3:7 to prepare piranha lotion, after the piranha lotion is cooled to room temperature, put the glass slide into the piranha lotion, heat to 90°C for 2 hours, take out the slide and wash with distilled water Wash 1-3 times; take a solution of 95% HCL and 99.7% absolute ethanol, mix it evenly at a ratio of 1:1 by volume, soak the slide in the solution and heat it to a constant temperature of 37°C, and then treat it with acid 1h, take out the slides and wash them with deionized water to complete the pretreatment of the slides, and store them in distilled water for later use; immerse the pretreated slides in the prepared 1% APTES anhydrous In ethanol solution, react at room tempera...

Embodiment 2

[0033] Cut the glass slide into small pieces of 10×10mm, soak in detergent for 5 minutes, rinse with tap water 1-3 times, then ultrasonically clean with deionized water for 5-10 minutes, and rinse with deionized water for 1-3 times; 30%H 2 o 2 , 98%H 2 SO 4Solution, according to the volume ratio of 3:7 to prepare piranha lotion, after the piranha lotion is cooled to room temperature, put the glass slide into the piranha lotion, heat to 90°C for 2 hours, take out the slide and wash with distilled water Wash 1-3 times; take a solution of 95% HCL and 99.7% absolute ethanol, mix it evenly at a ratio of 1:1 by volume, soak the slide in the solution and heat it to a constant temperature of 37°C, and then treat it with acid 1h, take out the slides and wash them with deionized water to complete the pretreatment of the slides, and store them in distilled water for later use; immerse the pretreated slides in the prepared 5% APTES anhydrous In ethanol solution, react at room temperat...

Embodiment 3

[0035] Cut the glass slide into small pieces of 10×10mm, soak in detergent for 5 minutes, rinse with tap water 1-3 times, then ultrasonically clean with deionized water for 5-10 minutes, and rinse with deionized water for 1-3 times; 30%H 2 o 2 , 98%H 2 SO 4 Solution, according to the volume ratio of 3:7 to prepare piranha lotion, after the piranha lotion is cooled to room temperature, put the glass slide into the piranha lotion, heat to 90°C for 2 hours, take out the slide and wash with distilled water Wash 1-3 times; take a solution of 95% HCL and 99.7% absolute ethanol, mix it evenly at a ratio of 1:1 by volume, soak the slide in the solution and heat it to a constant temperature of 37°C, and then treat it with acid 1h, take out the glass slide and wash it with deionized water, complete the pretreatment of the glass slide, and store it in distilled water for later use; immerse the pretreated glass slide in the prepared 10% APTES anhydrous In ethanol solution, react at ro...

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PUM

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Abstract

The invention discloses a method for fixing a DNA basic group to the surface of a glass slide. The method is suitable for fixing the DNA basic group to the surface of the glass slide. Firstly, the glass slide is cut into small pieces, washed, immersed in 95 percent of HCL and 99.7 percent of absolute ethyl alcohol mixed solution, processed through acid, and then is washed. Secondly, the glass slide is immersed in an APTES absolute ethyl alcohol to be prepared into a silanized glass slide. Thirdly, the silanized glass slide is immersed into a glutaraldehyde glutaraldehyde water solution to be reacted and flushed and is aired to be used later. Fourthly, a buffer solution is prepared, cosolvent and a surface active agent are added to the buffer solution to prepare a surface active agent solution, the DNA basic group is added to the surface active agent solution, full stirring is carried out to enable the DNA basic group to dissolve completely, and a DNA solution is prepared. Fifthly, the DNA solution is sampled to be dripped on the glass slide, the sampled DNA solution is cultivated in a calorstat at the appropriate temperature and is cleaned, and fixing of the DNA basic group is finished. By means of double modification of the glass slide, the wettability of the glass slide is improved, and the fixing rate of the DNA basic group is improved. Representation is carried out on the DNA basic group through a laser confocal raman spectrometer, the steps, needed in light splitting method of fluorescence, of fluorescence labeling and gene amplifying and hybridizing can be removed, and the DNA basic group fixing steps are greatly simplified.

Description

technical field [0001] The invention relates to a method for fixing DNA bases, in particular to a method suitable for fixing DNA bases on the surface of a glass slide. Background technique [0002] DNA is a helical structure composed of four bases: adenine (A, Adenine), guanine (G, Guanine), cytosine (C, Cytosine), and thymine (T, Thymine). The purine base is a double ring structure. The pyrimidine base is a single-ring structure. In the molecular structure of DNA, since the hydrogen bonds between the bases have a fixed number and the distance between the two DNA strands remains unchanged, the base pairing must follow certain rules, namely A=T, G≡C. This one-to-one correspondence between bases is called the principle of complementary base pairing. [0003] Nanopore-based third-generation gene sequencers or related key technologies are of strategic significance for protecting my country's own genetic resources. In order to realize gene sequencing technology, it is necessar...

Claims

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Application Information

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IPC IPC(8): G01N1/28
Inventor 罗勇刘秋真陈文文许浩
Owner CHINA UNIV OF MINING & TECH
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