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Nucleic acid fluorescence probe and preparation method thereof

A compound and alkyl technology, applied in the field of nucleic acid fluorescent probes and their preparation, can solve the problems of high cost, strong carcinogenicity and complex operation of nucleic acid probes

Inactive Publication Date: 2014-05-07
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional nucleic acid fluorescent probes include EB, YOYO, TOTO, Sybr Green, DAPI, and Hoechst dyes, which are used for the detection of double-stranded DNA with high sensitivity, but the detection effect for single-stranded DNA, especially short-chain single-stranded DNA very poor
At present, there are no fluorescent dyes sensitive enough to single-stranded DNA to replace fluorescent-labeled or isotope-labeled DNA probes, especially in the application of nucleic acid electrophoresis, fluorescent-labeled or isotope-labeled nucleic acid probes are expensive and complicated to operate
In addition, among the traditional nucleic acid dyes, the intercalating agent represented by EB has strong carcinogenicity; while the dyes with low carcinogenicity, such as the Sybr series, have a complicated synthesis process, resulting in high prices, and their application is far less extensive than that of EB.

Method used

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  • Nucleic acid fluorescence probe and preparation method thereof
  • Nucleic acid fluorescence probe and preparation method thereof
  • Nucleic acid fluorescence probe and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The synthesis of embodiment 1 probe 4 and 5

[0035]

[0036] Mix 0.82g of a1 (synthesized by reference Chem.Commun.2006, 3705–3707), 0.24g of sodium azide and 15mL of dimethyl sulfoxide, stir at 80°C for 3 hours, pour into water, and the precipitated insoluble Purified by column chromatography to obtain 0.7 g of light yellow oil, ie intermediate b, with a yield of 98%. 1 H NMR (300MHz, CDCl 3 )δ[ppm]:7.15-7.0(m,10H),7.00-6.90(m,4H),6.69-6.62(m,4H),4.11-4.04(m,4H),3.59-3.52(m,4H) .

[0037]Dissolve 0.5g of b and 1.07g of triphenylphosphine in 120mL of tetrahydrofuran, add 20mL of water, stir overnight at 60°C, distill off the solvent under reduced pressure, and use chloroform-methanol gradient eluent column chromatography to obtain probe 5 in sequence. and 4 were concentrated under reduced pressure to obtain 0.19 g and 0.185 g of white solids, and the yields were 44% and 41%, respectively. Probe 4: ESI-MS m / z[M+H] + 451; Probe 5: ESI-MS m / z [M+H] + 451.

Embodiment 2

[0038] The synthesis of embodiment 2 probe 8

[0039]

[0040] Using a2 (synthesized in reference Chem.Eur.J.2008,14,6428–6437) as raw material, according to the synthesis method of probe 4 and probe 5, the amount of sodium azide and triphenylphosphine was doubled, and the Probe 8 was obtained as a white solid. ESI-MS m / z[M+H] + 569.

Embodiment 3

[0041] Synthesis of Example 3 Probes 11 and 12

[0042]

[0043] Using a3 (synthesized by reference Org. Lett. 2008, 10, 4581–4584) as raw material, according to the synthesis method of probe 4 and probe 5, probes 11 and 12 can be obtained as white or yellow solids. Probe 11: ESI-MSm / z[M+H] + 507; Probe 12: ESI-MS m / z [M+H] + 507.

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Abstract

The invention discloses a novel nucleic acid fluorescence probe. Amido is introduced into tetraphenyl ethylene, amido is a basic group and can be combined with nucleic acid, so that molecular rotation is limited, nonradiative excited-state relaxation is inhibited and fluorescence is obviously enhanced, and thus nucleic acid detection is realized. The novel nucleic acid fluorescence probe has high sensitivity to double-stranded DNA and single-stranded DNA, and the probe has good application prospect to gel electrophoresis or detection of nucleic acid in biological samples.

Description

technical field [0001] The invention relates to a nucleic acid fluorescent probe and a preparation method thereof, belonging to the technical field of biological detection. Background technique [0002] Nucleic acid is the genetic material of life and plays an important role in life activities. Nucleic acid detection has important applications in the fields of disease diagnosis, environmental microbial identification, and aptamers. Therefore, the development of fluorescent detection probes for nucleic acids is of great significance. Traditional nucleic acid fluorescent probes include EB, YOYO, TOTO, Sybr Green, DAPI, and Hoechst dyes, which are used for the detection of double-stranded DNA with high sensitivity, but the detection effect for single-stranded DNA, especially short-chain single-stranded DNA very bad. At present, there are no fluorescent dyes sensitive enough to single-stranded DNA to replace fluorescent-labeled or isotope-labeled DNA probes, especially in the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C209/42C07C211/27C07C213/02C07C217/18C12Q1/68
Inventor 杨楚罗朱策泽徐黎
Owner WUHAN UNIV
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