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Hairpin DNA template of fluorescent nano copper clusters, and applications thereof

A fluorescent nanometer and DNA sequence technology, which is applied in the direction of recombinant DNA technology, DNA/RNA fragments, microbial measurement/inspection, etc., to achieve the effects of rapid response, mild formation conditions, and high fluorescence intensity

Inactive Publication Date: 2017-01-04
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, for now, the research on fluorescent nano-copper clusters is still in its infancy. It is necessary to develop new templates for fluorescent nano-copper clusters, especially new DNA templates, and improve the fluorescence properties of fluorescent copper clusters to further broaden the scope of fluorescent copper nano-clusters. The scope of application is an important direction for the development of this field

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  • Hairpin DNA template of fluorescent nano copper clusters, and applications thereof
  • Hairpin DNA template of fluorescent nano copper clusters, and applications thereof
  • Hairpin DNA template of fluorescent nano copper clusters, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Hairpin DNA templates form fluorescent nano-copper clusters and compare experiments with linear DNA templates

[0044] The DNA sequences used are: hairpin DNA templates with loop T numbers of 6, 10, 15, 20, 30 and 40 respectively (such as figure 1 —DNA 1-5), the stems all adopt RS1 double strands with weak ability to form fluorescent copper clusters, in addition, single-stranded linear poly T is used as a control, including T6, T10, T20, T30 and T40 (such as figure 1 — DNA6-10);

[0045] Schematic diagram of the formation of fluorescent copper clusters based on linear double-stranded templates, poly T templates and MB-Tn templates. figure 2 shown;

[0046] Take 100pmol of DNA 1-10 respectively, put them into 1.5mL EP tubes, add MOPs buffer (10mM OPs, 150mM NaCl, pH 7.0) to 194μL, then add 2μL of 10mM copper sulfate solution, 4μL of 100mM vitamin C solution, React in a shaker at room temperature for 10 minutes;

[0047] Fluorescence measurement, the excit...

Embodiment 2

[0049] Example 2: Effect experiment of the ring base of the hairpin DNA template on the formation of fluorescent nano-copper clusters

[0050] The DNA sequence used is: the hairpin DNA with T20, A20, C20 and (CT)10 in the loop, and the RS1 double strand in the stem (such as figure 1 — DNA 3, 11-13);

[0051] Take 100 pmol of the above hairpin sequence into 1.5 mL EP tubes, dilute to 194 μL with MOPs buffer, add 2 μL of 10 mM copper sulfate solution, 4 μL of 100 mM vitamin C solution, and react in a shaker at room temperature for 10 minutes;

[0052] Fluorescence measurement, the excitation wavelength is 345nm, and the emission wavelength scanning range is 500nm-650nm;

[0053] The results show that the fluorescence spectrum of the fluorescent copper nanoclusters formed by the hairpin template is as attached Figure 5 As shown, a strong fluorescent signal can be formed only when the T base is used as the loop.

Embodiment 3

[0054] Example 3: Effect of the length of the stem sequence on the formation of fluorescent copper clusters

[0055] The DNA sequences used are: the loops are all T20, the stems are GC double-stranded sequences of different lengths, hairpin DNA templates with lengths of 6, 8, 10, 12, 14, 16, and 18 bp respectively (such as figure 1 —DNA 14-20),

[0056] Take 100 pmol of the above hairpin sequence into 1.5 mL EP tubes, dilute to 194 μL with MOPs buffer, add 2 μL of 10 mM copper sulfate solution, 4 μL of 100 mM vitamin C solution, react in a shaker at room temperature for 10 minutes,

[0057] Fluorescence measurement, excitation wavelength is 345nm, emission wavelength scanning range is 500nm-650nm,

[0058] The fluorescence spectra of the fluorescent copper nanoclusters formed by the prepared hairpin template are as follows: Figure 6 As shown, when the base pair of the stem sequence is between 6-12, the fluorescence intensity increases with the extension of the length of the...

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Abstract

The invention belongs to the technical field of functional material and fluorescent nano probes, and relates to a novel hairpin DNA template used for synthesis of fluorescent nano copper clusters, and applications of the novel hairpin DNA template in biomolecule detection. The novel hairpin DNA template possesses a hairpin structure; a loop part is composed of poly T with different length, and a stem part is composed of double-stranded DNA with different length and base sequences; and in a certain buffer solution, fluorescence signals can be generated via a few minutes of reaction at room temperature under the action of copper sulfate and vitamin C. Compares with linear poly T, the novel hairpin DNA template possesses following advantages: the fluorescence intensity of the formed fluorescent nano copper clusters is higher, and possesses poly T length and sequence concentration dependence; formation of the fluorescent nano copper clusters is influenced by the stability of the novel hairpin DNA template; formation conditions are mild; reaction is quick; fluorescence intensity is higher than that of the prior art; and biomolecule detection technology without labeling or separating can be designed easily.

Description

technical field [0001] The invention relates to the field of functional materials and the technical fields of fluorescent nano-probes and molecular recognition and biosensing, and specifically discloses a novel DNA template for synthesizing fluorescent nano-copper clusters and its application in biomolecular detection. Background technique [0002] The prior art discloses that fluorescence analysis technology has simple and intuitive characteristics, as well as time and space resolution capabilities, and this technology is widely used in the detection of target molecules in vivo and in vitro [Wu J., Liu W., Ge J., Zhang H., Wang P. Chem. Soc. Rev., 2011, 40, 3483–3495]. Typical ones include organic fluorescent dye molecules [Dsouza R.N., Pischel U., Nau W.M., Chem. Rev., 2011, 111(12), 7941-7980], aggregation-induced enhancement and quenching fluorescent dyes [Wang B., Yu C.,Angew.Chem.Int.Ed.,2010,49(8):1485-1488; Hu Q.,Gao M.,Feng G.,Chen X.,Liu B.ACS Appl.Mater.Interface...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 曹志娟王亦男卢建忠崔红艳刘彩云
Owner FUDAN UNIV
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