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Polythymine template, fluorescent copper nano-cluster based on same, preparation method of fluorescent copper nano-cluster and ATP detection method

A technology of fluorescent copper nanoclusters and polythymine, which is applied in biochemical equipment and methods, DNA preparation, and microbial determination/inspection, etc., can solve the problems of complicated steps, unsuitable for rapid detection, high cost, etc., and achieves easy operation. , the detection steps are simple and easy, and the cost is low.

Inactive Publication Date: 2015-11-25
SHENZHEN INST OF ADVANCED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although its detection accuracy is high, the cost is high, and the steps are relatively complicated, so it is not suitable for rapid detection

Method used

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  • Polythymine template, fluorescent copper nano-cluster based on same, preparation method of fluorescent copper nano-cluster and ATP detection method
  • Polythymine template, fluorescent copper nano-cluster based on same, preparation method of fluorescent copper nano-cluster and ATP detection method
  • Polythymine template, fluorescent copper nano-cluster based on same, preparation method of fluorescent copper nano-cluster and ATP detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0151] 1) Provide polythymine template

[0152] The nucleotide sequences of synthetic oligonucleotides T25, A25, T25-biotin, T25-a1, T25-b1, T25-b2, T25-C10-T25 and T25-C10-T25-C10-T25 are shown in Table 1 below Shown:

[0153]

[0154]

[0155] Table 1. Oligonucleotides T25, A25, T25-biotin, T25-a1, T25-b1,

[0156] Nucleotide sequences of T25-b2, T25-C10-T25 and T25-C10-T25-C10-T25

[0157] Oligonucleotides T25, A25, T25-biotin, T25-a1, T25-b1, T25-b2, T25-C10-T25 and T25-C10-T25-C10-T25 as described in Table 1 are single Stranded deoxyribonucleic acid. Wherein, T25, T25-biotin, T25-a1, T25-b1, T25-b2, T25-C10-T25 or T25-C10-T25-C10-T25 are polythymine templates.

[0158] 2) Configure reagents

[0159] a. Configuration of polythymine template solution: Take the T25 single-stranded polythymine sequence described in step (1), dilute 10OD T25 with 510 μL ultrapure water to 100 μM, and set aside; meanwhile, follow the same laboratory routine operation Prepare A25, T2...

Embodiment 2

[0166] This embodiment provides a method for preparing fluorescent copper nanoclusters based on a polythymine template, comprising the following steps:

[0167] (1) Configure MOPS buffer, 100 μM T25, ascorbic acid and copper sulfate solution according to the operation described in Example 1 step (2);

[0168] (2) Add 192 μL of MOPS buffer, 1 μL of LT25, 5 μL of ascorbic acid, and 2 μL of copper sulfate solution in sequence in the EP tube, and shake gently for 5 minutes to obtain the fluorescent copper nanocluster based on the polythymine template. The fluorescent copper The fluorescence of the nanoclusters is red, wherein the ascorbic acid is in excess, and the molar ratio of T25 to copper sulfate is 1:200.

[0169] In order to fully illustrate the beneficial effects of the method for preparing fluorescent copper nanoclusters based on polythymine templates provided by the present invention, this embodiment also provides fluorescence spectral data of the prepared fluorescent co...

Embodiment 3

[0196] This embodiment provides a method for preparing fluorescent copper nanoclusters based on a polythymine template, comprising the following steps:

[0197] (1) Configure MOPS buffer, 100 μM T25-C10-T25, ascorbic acid and copper sulfate solution according to the operation described in step (2) of Example 1;

[0198](2) Add 192 μL of MOPS buffer, 1 μL of LT25, 5 μL of ascorbic acid, and 2 μL of copper sulfate solution in sequence in the EP tube, and shake gently for 5 minutes to obtain the fluorescent copper nanocluster based on the polythymine template. The fluorescent copper The fluorescence of the nanoclusters is red, wherein the ascorbic acid is in excess, and the molar ratio of T25 to copper sulfate is 1:200.

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Abstract

The invention provides a polythymine template, a fluorescent copper nano-cluster based on the same, a preparation method of the fluorescent copper nano-cluster and an ATP detection method. The polythymine template has a single-chain DNA sequence and comprises at least one polythymine sequence which is (T)n, wherein n refers to natural number ranging from 15 to 40. The polythymine template can be used for preparing the fluorescent copper nano-cluster, preparation cost is low, and operation is simple and convenient. The ATP detection method is low in cost, simple and easy-to-implement in detection step, obvious in phenomenon, easy for observation and capable of quickly detection concentration of ATP in a sample in a high-sensitivity manner.

Description

technical field [0001] The invention relates to the field of nanomaterials and molecular detection, in particular to a polythymine template, a fluorescent copper nanocluster based on the polythymine template and a preparation method thereof, and an ATP detection method. Background technique [0002] ATP has an important influence on the activity of many enzymes, and its increase or decrease will cause various diseases, such as hypoxia, hypoglycemia, Parkinson's disease and malignant tumors. Therefore, simple and rapid detection of ATP is crucial for the treatment of related diseases. The current method for detecting ATP mainly uses luciferin and luciferase detection. Although its detection accuracy is high, the cost is high and the steps are relatively complicated, so it is not suitable for rapid detection. [0003] Therefore, it is necessary to provide a low-cost, easy-to-operate method that can detect ATP quickly and with high sensitivity. Contents of the invention ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12N15/10B22F9/24
Inventor 蔡林涛吴亚运张鹏飞高笃阳陈驰
Owner SHENZHEN INST OF ADVANCED TECH
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