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Novel use of cyanine dye in detection of G-quadruplex DNA

A technology of cyanine dyes and quadruplexes, which is applied in the field of new applications of cyanine dyes in the detection of G-quadruplex structure DNA, which can solve the problems of high price, difficult separation and purification, and high requirements for instruments, so as to overcome the long period and cost cheap effect

Inactive Publication Date: 2011-03-30
INST OF CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Novel use of cyanine dye in detection of G-quadruplex DNA
  • Novel use of cyanine dye in detection of G-quadruplex DNA
  • Novel use of cyanine dye in detection of G-quadruplex DNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Embodiment 1, the detection of G-quadruplex structure DNA

[0066] 1. Preparation of reaction solution

[0067] 1) Solution A

[0068] 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.

[0069] 2) Solution B

[0070] 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.

[0071] 3) Solution C

[0072] Add 40 μL of 200.0 μM methanol (excellent grade) solution of cyanine dye to a 2 ml volumetric flask, dilute to volume with pho...

Embodiment 2

[0087] Embodiment 2, the detection of G-quadruplex structure DNA

[0088] 1. Preparation of reaction solution

[0089] 1) Solution A

[0090] Add 40 μL of 200.0 μM methanol solution of cyanine dye to a 2 ml volumetric flask; add 20 μL of 200 μM phosphate buffer solution (pH 6.0) of sample 1, dilute to volume with phosphate buffer solution (pH 6.0), and mix to obtain solution A. In solution A, the molar ratio of sample 1 DNA to cyanine dye is 0.5:1.

[0091] 2) Solution B

[0092] Add 40 μL of 200.0 μM methanol solution of cyanine dye to a 2 ml volumetric flask; add 20 μL of 200 μM phosphate buffer solution (pH 6.0) of sample 2, dilute to volume with phosphate buffer solution (pH 6.0), and mix to obtain solution B. In solution B, the molar ratio of sample 2 DNA to cyanine dye is 0.5:1.

[0093] 3) Solution C

[0094] Add 40 μL of a 200.0 μM methanol solution of cyanine dye molecules into a 2 ml volumetric flask, dilute to the volume with phosphate buffer (pH 6.0), and mix ...

Embodiment 3

[0106] Embodiment 3, the detection of G-quadruplex structure DNA

[0107] 1. Preparation of reaction solution

[0108] 1) Solution A

[0109] Add 40 μL of 200.0 μM pure methanol solution of cyanine dye to a 2ml volumetric flask; add 160 μL of 200 μM phosphate buffer solution (pH 8.0) of sample 1, dilute to volume with phosphate buffer solution (pH 6.0), and mix to obtain solution A. In solution A, the molar ratio of sample 1 DNA to cyanine dye is 4:1.

[0110] 2) Solution B

[0111] Add 40 μL of 200.0 μM pure methanol solution of cyanine dye to a 2 ml volumetric flask; add 160 μL of 200 μM phosphate buffer solution (pH 8.0) of sample 2, dilute to volume with phosphate buffer solution (pH 6.0), and mix to obtain solution B. In solution B, the molar ratio of sample 2 DNA to cyanine dye is 4:1.

[0112] 3) Solution C

[0113] Add 40 μL of 200.0 μM pure methanol solution of cyanine dye molecules into a 2 ml volumetric flask, dilute to the volume with phosphate buffer (pH 8.0)...

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Abstract

The invention provides novel use of a cyanine dye in the detection of a G-quadruplex DNA. The detection of the G-quadruplex DNA by using the cyanine dye has the advantages of simplicity, quickness andrelatively low prices and overcomes the drawbacks of the prior detection technology such as long period, high price and high technical and equipment requirements. The use of the cyanine dye in the detection of the G-quadruplex DNA makes whether a DNA sample in solution has a G-quadruplex structure or a linear double helical structure determined quickly by a UV-visible absorption spectrum, fluorescence spectrum or confocal laser scanning microscopy. By using confocal laser scanning microscopy, the DNA sample assembled on the surface of Au can be marked visually, so the structure of the DNA sample can be recognized and the specific assembly position of the DNA sample on the surface of the Au can be marked at the same time.

Description

technical field [0001] The invention relates to a new application of cyanine dye in detecting DNA with G-quadruplex structure. 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 controve...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/33C12Q1/68
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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