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DNA liquid crystal biosensor and its preparation method and detection method

A biosensor and DNA probe technology, applied in the field of biosensors, can solve problems such as the influence of DNA probe hybridization efficiency, influence on sensor repeatability, and increase sensor manufacturing costs, achieving good selectivity and sensitivity, low cost, and easy production easy effect

Inactive Publication Date: 2020-12-15
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above group modification for DNA not only increases the cost of sensor fabrication, but also the hybridization efficiency of DNA probes is also affected by the modified groups on it.
At the same time, this kind of fixing method has many steps, which will affect the repeatability of the sensor

Method used

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  • DNA liquid crystal biosensor and its preparation method and detection method
  • DNA liquid crystal biosensor and its preparation method and detection method
  • DNA liquid crystal biosensor and its preparation method and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: the preparation method and detection of a kind of DNA liquid crystal biosensor:

[0030] (1) Gold-plated glass slides

[0031] 1.1. Cleaning slides: ① Soak 12mm×12mm slides in 70% H by volume 2 SO 4 : 30%H 2 o2 In the prepared piranha solution, react at 80°C for 1 h; ②Rinse 5 times with ultrapure water; ③Sonicate in ethanol for 10 min; ④Repeat ②, then sonicate in methanol for 10 min; Dry in a nitrogen atmosphere for use;

[0032] 1.2, the surface of the glass slide is plated with a uniform gold film with a nano-scale thickness by vacuum evaporation to obtain a gold-plated base; the gold film is composed of a 3nm chromium adhesion layer and a 10nm gold film layer, and the prepared gold-plated glass The film has good light transmittance; the gold-plated substrate is cleaned according to the following steps: ① Rinse the gold-plated substrate 5 times with ultrapure water; ② Ultrasonic in ethanol for 10 min; Dry; ④Put the gold-plated substrate in a UV clea...

Embodiment 2

[0044] Embodiment 2: Sensor Background Noise Control

[0045] In the scheme of this application, the orientation of the liquid crystal molecules in the liquid crystal cell is determined by the surface of the working substrate, more specifically, the orientation of the liquid crystal molecules is mainly affected by the immobilization density of the capture probes on the substrate: when the capture probe immobilization density When the density of the capture probes is too high, the liquid crystal molecules can not be embedded in the space between the single-stranded DNA. As a result, the single-stranded DNA at this time cannot induce the vertical alignment of liquid crystals. In order to investigate the background noise of the novel DNA liquid crystal biosensor, by adjusting the A in the DNA probe fixative solution in the DNA biosensor preparation process described in Example 1 m P 1 with A n The preferred value of y, m and n is 15, and a series of working film substrates fix...

Embodiment 3

[0046] Embodiment 3: target DNA sequence detection

[0047] A novel DNA liquid crystal biosensor is used to detect the target DNA sequence. Prepare a series of working substrates with the preferred parameters m and n preferably set at 15, y = 7:5, and drop test solutions with different target DNA concentrations on the substrates to carry out molecular recognition reactions, and assemble according to the aforementioned steps The liquid crystal cell was placed under a polarizing microscope to collect images. The result is as image 3 shown. It can be seen that with the increase of the concentration of target DNA in the liquid to be detected, the image has a color change of black-gray-yellow-pink-green in sequence. This is because when the target DNA concentration in the liquid to be tested is greater, the more target DNA is bound to the working substrate, the greater the disturbance to the liquid crystal orientation on the substrate, and the greater the inclination angle of t...

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Abstract

The invention discloses a DNA liquid crystal biological sensor. The DNA liquid crystal biological sensor comprises a working substrate prepared by jointly fixing a capture probe and an isolation probeon a slide with the surface being plated with a layer of nanoscale-thickness gold film, wherein the capture probe consists of a section of base sequence P containing m adenine and is specifically bonded to a to-be-detected object and is marked as AmP1; the isolation probe contains poly-adenine of n adenine A and is marked as An, m is a positive integer, and n is a natural number. The invention also discloses a preparation method and a detection method of the DNA liquid crystal biological sensor. The DNA liquid crystal biological sensor obtained in the invention has the same advantages with aliquid crystal sensor, the probe can efficiently identify the to-be-detected object without any chemical modification, and the preparation procedures are simplified; the production cost is reduced; and the DNA liquid crystal biological sensor can also be applied to the biological molecular detection according to nucleic acid hybridization and aptamer molecular identification mechanism.

Description

technical field [0001] The invention belongs to the technical field of biosensors, and in particular relates to a DNA liquid crystal biosensor, a preparation method and a detection method thereof. Background technique [0002] Deoxyribonucleic acid (DNA), a biological macromolecule containing genetic information, controls the development of organisms and the operation of vital functions, and can directly affect human genetics and health. Therefore, the detection of DNA plays an extremely important role in many fields such as: clinical diagnosis, medical research, agricultural production, food safety, environmental monitoring, etc. [0003] Traditional DNA detection techniques include: PCR, chemiluminescence analysis, electrophoresis related techniques, etc. However, these techniques are cumbersome to operate, have many steps, and must be equipped with expensive professional equipment or professionals, making it difficult to perform in-situ and real-time detection far from t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/84C12Q1/6825
CPCC12Q1/6825G01N21/84
Inventor 熊兴良刘慧龙陈龙聪江奇锋梁波
Owner CHONGQING MEDICAL UNIVERSITY
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