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A fluorescence detection method that distinguishes single and dual -chain nucleotide

A new method, fluorescence analysis technology, applied in biochemical equipment and methods, fluorescence/phosphorescence, microbial determination/inspection, etc., can solve the problems of unfavorable π-electron interaction, small conjugated system, etc., to achieve low detection cost, Controllable effects of the preparation process

Inactive Publication Date: 2011-12-28
QINGDAO UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most of these sensing systems require expensive fluorescently-labeled DNA probes and the conjugated system of polymeric units is small, which is not conducive to the generation of π-electron interactions between molecular chains.
In the known literature, there is no report on the use of polymers containing fused ring structures to detect and distinguish single-stranded (ssDNA) and double-stranded nucleotides (dsDNA) through the fluorescence recovery of the sensing system.

Method used

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  • A fluorescence detection method that distinguishes single and dual -chain nucleotide
  • A fluorescence detection method that distinguishes single and dual -chain nucleotide

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Experimental program
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Effect test

Embodiment 1

[0024] Weigh an appropriate amount of the synthesized conjugated oligopyrene derivative and place it in a 15mL reagent bottle. Prepare a concentration of 10mmolL with ultrapure water -1 ; Tris-HCl buffer pH=7.4. Afterwards, add a certain volume of buffer solution into the reagent bottle containing the conjugated oligopyrene derivative to make the concentration 3.34×10 -6 mol L -1 , and pipette 3 parts of 5mL of the solution as the detection solution. Then, drop the ssDNA and dsDNA samples to be tested into the system. The final concentrations of ssDNA and dsDNA were both 8×10 -7 mol L -1 . The fluorescence response of the conjugated oligopyrene derivative detection solution to DNA is obtained by recording the fluorescence intensity of the fluorescence sensing system at 465 nm with a FL 4600 fluorescence instrument at 25 ° C, and the excitation wavelength is 340 nm. The experimental results are: ssDNA and dsDNA make the recovery values ​​of the fluorescent signals of the...

Embodiment 2

[0026] Weigh an appropriate amount of the synthesized conjugated oligopyrene derivative and place it in a 15mL reagent bottle. Prepare a concentration of 10mmolL with ultrapure water -1 ; Tris-HCl buffer pH=7.4. Afterwards, add a certain volume of buffer solution into the reagent bottle containing the conjugated oligopyrene derivative to make the concentration 3.34×10 -6 mol L -1 , and pipette 3 parts of 5mL of the solution as the detection solution. Then, drop the ssDNA and dsDNA samples to be tested into the system. The final concentrations of ssDNA and dsDNA were both 3×10 -5 mol L -1 . The fluorescence response of the conjugated oligopyrene derivative detection solution to DNA is obtained by recording the fluorescence intensity of the fluorescence sensing system at 465 nm with a FL 4600 fluorescence instrument at 25 ° C, and the excitation wavelength is 340 nm. The experimental results are: ssDNA and dsDNA make the recovery values ​​of the fluorescent signals of the...

Embodiment 3

[0028] Weigh an appropriate amount of the synthesized conjugated oligopyrene derivative and place it in a 15mL reagent bottle. Prepare a concentration of 10mmolL with ultrapure water -1 ; Tris-HCl buffer pH=7.4. Afterwards, add a certain volume of buffer solution into the reagent bottle containing the conjugated oligopyrene derivative to make the concentration 3.34×10 -6 mol L -1 , and pipette 3 parts of 5mL of the solution as the detection solution. Then, drop the ssDNA and dsDNA samples to be tested into the system. The final concentrations of ssDNA and dsDNA were both 8×10 -5 mol L -1 . The fluorescence response of the conjugated oligopyrene derivative detection solution to DNA is obtained by recording the fluorescence intensity of the fluorescence sensing system at 465nm with a FL 4600 fluorescence instrument at 25°C, and the excitation wavelength is 340nm. The experimental results are: ssDNA and dsDNA make the recovery values ​​of the fluorescent signals of the con...

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Abstract

A new fluorescence analysis method for distinguishing ssDNA and dsDNA based on oligopyrene derivatives, which includes the following specific steps: (1) Synthesizing water-soluble oligopyrene derivatives by chemical methods and electrochemical techniques, the synthesis route is relatively simple, and the product Purification is easier; (2) The fluorescent conjugated oligopyrene derivative is dissolved in Tris-HCl buffer to prepare a solution with a certain concentration. Based on the difference in recovery of the fluorescent signal of the conjugated oligopyrene derivatives induced by ssDNA and dsDNA, it is convenient to distinguish ssDNA and dsDNA. Different from many existing methods for detecting DNA based on conjugated polymers, the detection system involved in the present invention does not contain expensive fluorescent-labeled DNA probes, so the detection cost is low and has good practical application prospects.

Description

technical field [0001] The invention relates to a fluorescence detection method for distinguishing single-stranded (ssDNA) and double-stranded nucleotides (dsDNA), belonging to the technical field of material preparation. technical background [0002] In the study of biosensing systems, the detection and analysis of DNA, proteins, enzymes, and small biological molecules are very important research contents. Among them, the quantitative analysis and specific identification of DNA and RNA are of great significance in the fields of genomics, virology, molecular biology, disease diagnosis, gene mutation and gene transfer. However, there are still great difficulties and challenges in the rapid, sensitive, and low-cost detection and identification of DNA, and these technologies are the key link in the practical application of DNA detection and analysis. [0003] Recent research work at home and abroad has proved that the sensitivity of fluorescent conjugated polymers is much high...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64
Inventor 陈友强张新霓陈利祥
Owner QINGDAO UNIV
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