Application of supramolecular ordered structure in detection of G-quadruplex DNA

A supramolecular aggregate and supramolecular technology, which is applied in the application field of supramolecular ordered body in the detection of G-quadruplex structure DNA, can solve the problems of difficult preservation of protein activity, high equipment requirements, high price and the like.

Inactive Publication Date: 2009-11-25
INST OF CHEM CHINESE ACAD OF SCI
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Problems solved by technology

However, it is difficult to separate and purify the protein with high purity as the detection object. In addition, the protein activity is not easy to preserve, and the price is also very expensive, which greatly limits the scope of application of the above methods.
It has been reported that a single fluorescent molecule is used to specifically label G-quadruplex structure DNA, but this method has complicated detection methods and very high requirements for instruments, and basically cannot be widely used

Method used

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  • Application of supramolecular ordered structure in detection of G-quadruplex DNA
  • Application of supramolecular ordered structure in detection of G-quadruplex DNA
  • Application of supramolecular ordered structure in detection of G-quadruplex DNA

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Embodiment 1

[0028] Embodiment 1, application of cyanine dye supramolecular ordered body to detect G-quadruplex structure DNA

[0029] 1. Preparation of reaction solution

[0030] 1) Solution A

[0031] Add 40 μL 200.0 μM methanol (excellent grade) solution of cyanine dye to a 2ml volumetric flask; add 60 μL 200 μM phosphate buffer solution (pH 6.0) of sample 1, dilute to volume with phosphate buffer solution (pH 6.0), mix well, Solution A is obtained. In solution A, the molar ratio of sample 1 DNA to cyanine dye is 1.5:1.

[0032] 2) Solution B

[0033] Add 40 μL 200.0 μM methanol (excellent grade) solution of cyanine dye to a 2ml volumetric flask; add 60 μL 200 μM phosphate buffer solution (pH 6.0) of sample 2, dilute to volume with phosphate buffer solution (pH 6.0), mix well, Solution B is obtained. In solution B, the molar ratio of sample 2 DNA to cyanine dye is 1.5:1.

[0034] 3) Solution C

[0035] Add 40 μL of 200.0 μM methanol (excellent grade) solution of cyanine dye to a ...

Embodiment 2

[0050] Embodiment 2, application of cyanine dye supramolecular order body to detect G-quadruplex structure DNA

[0051] 1) Respectively react sample 3 and sample 4 with Au sheets treated with mercaptoundecanoic acid to assemble DNA onto the surface of Au sheets respectively to obtain Au sheet A and Au sheet B;

[0052] 2) Put the processed Au sheet into a cyanine dye phosphate buffer solution (pH 6.0) with a concentration of 10 μM and dye it in the dark for 1 hour at room temperature;

[0053] 3) After taking out the Au sheet, rinse it repeatedly with phosphate buffer to wash off the cyanine dye aggregates directly adsorbed on the surface of the Au sheet;

[0054] 4) After blowing dry with nitrogen, place the Au sheet under a confocal laser scanning microscope (CLSM), use a 559nm laser to excite the fluorescence of the cyanine dye molecules, and receive the emission fluorescence images of the cyanine dye molecules by wavelength.

[0055] See Figure 7 , from the CLSM fluores...

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Abstract

The invention discloses the use of a supramolecular ordered structure in the detection of a G-quadruplex DNA. The method realizes the detection of the G-quadruplex DNA by detecting the change of the aggregation state of the supramolecular ordered structure. The detection of the G-quadruplex DNA by using the supramolecular ordered structure has the advantages of simplicity, quickness and relatively low price and overcomes the drawbacks of the prior detection technology such as long period, high price and high technical and equipment requirements.

Description

technical field [0001] The invention relates to the application of supramolecular ordered body in detecting G-quadruplex structure DNA. Background technique [0002] Single-stranded telomeric DNA easily undergoes four base pairs through hydrogen bonds between guanine bases, forming a planar G-quadruplex structure. Human Telomere DNA Sequence d(TTAGGG) 4 G-quadruplexes of different structures can be formed under the action of potassium ions or sodium ions. Identify G-quadruplex DNA in in vitro and in vivo experiments (mainly different from linear double-helix DNA), which is useful for determining the physiological functions of G-quadruplex DNA in the human body and the development of anti-tumor drugs, etc. aspects are very important. [0003] At present, there have been some reports in the literature on the recognition of G-quadruplex structure DNA in vitro and in vivo. It is difficult to identify G-quadruplex structure DNA in vivo experiments, and there is still a lot of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N21/33C12Q1/68
Inventor 唐亚林杨千帆向俊锋徐广智
Owner INST OF CHEM CHINESE ACAD OF SCI
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