DNA nanosphere detection probe as well as preparation method and application thereof

A technology for detecting probes and nanospheres, applied in the field of biosensing, can solve problems such as poor results and achieve low detection limits

Pending Publication Date: 2022-01-28
CHANGSHA UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention provides a DNA nanosphere detection probe in order to overcome the defects that the above-mentioned existing detection methods are not effective.

Method used

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  • DNA nanosphere detection probe as well as preparation method and application thereof
  • DNA nanosphere detection probe as well as preparation method and application thereof
  • DNA nanosphere detection probe as well as preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0044] Embodiment 1 provides a DNA nanosphere detection probe, and the specific preparation method includes the following steps:

[0045] First, under the action of telomerase, the single-stranded probe C is extended to obtain the DNA telomerase product chain D-STP;

[0046] S11. Mix 10 μL DNA telomerase product chain D-STP with single-stranded probe A, and react at room temperature for 30 minutes to obtain a mixed solution, the concentration of the single-stranded probe A is 75 nM;

[0047] S12. Add the single-stranded probe B to the mixture in step S11 and react at room temperature for 180 minutes; the concentration of the single-stranded probe B is 75 nM to obtain a DNA nanosphere detection probe.

[0048] The single-stranded probe A is shown in SEQ ID No.1 in the sequence listing, the single-stranded probe B is shown in SEQ ID No.4 in the sequence listing, and the nucleotide of the single-stranded probe C is The sequence is shown as SEQ ID No.7 in the sequence listing.

Embodiment 2

[0050] The preparation method of embodiment 2 is the same as that of embodiment 1, and the only difference is that the single-stranded probe A is as shown in SEQ ID No.2 in the sequence listing, and the single-stranded probe B is as SEQ ID No. in the sequence listing. 5, the nucleotide sequence of the single-stranded probe C is shown in SEQ ID No.8 in the sequence listing.

Embodiment 3

[0052] The preparation method of embodiment 3 is the same as that of embodiment 1, and the only difference is that the single-stranded probe A is as shown in SEQ ID No.3 in the sequence listing, and the single-stranded probe B is as shown in SEQ ID No.3 in the sequence listing. 6, the nucleotide sequence of the single-stranded probe C is shown in SEQ ID No.9 in the sequence listing.

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Abstract

The invention discloses a DNA nanosphere detection probe as well as a preparation method and an application thereof. The DNA nanosphere detection probe comprises a single-stranded probe A, a single-stranded probe B and a single-stranded probe C, each of the single-stranded probe A, the single-stranded probe B and the single-stranded probe C at least contains a cohesive end of a section of palindromic sequence, each section of palindromic sequence comprises 10 basic groups, the single-stranded probe C further comprises a primer sequence for telomerase reaction, telomere repetitive sequences of the single-stranded probe A, the single-stranded probe B and a DNA telomerase product chain D-STP are partially complementary to form DNA of a Y-type structure, the DNA of the Y-type structure is connected by a viscous end and is sequentially amplified and arranged to obtain the DNA nanosphere detection probe, and the diameter of the DNA nanosphere detection probe is 10-100 nm. The DNA nanosphere detection probe can accurately detect the activity of telomerase in different cell lines, and has the advantages of low sensitivity, low detection limit, rapid recognition and the like.

Description

technical field [0001] The invention relates to the technical field of biosensing, and more specifically, to a DNA nanosphere detection probe and its preparation method and application. Background technique [0002] A telomere is a DNA sequence with a repeating sequence (TTAGGG) that usually exists at the end of a cell's chromosomes. During cell division, the duplication of chromosomes leads to the continuous shortening of telomeres. As a reverse transcriptase widely present in the body, telomerase plays a key role in maintaining telomere length and in the process of cell growth and differentiation. Telomerase achieves the purpose of maintaining telomere length and cell activity by continuously replicating the TTAGGG repeat sequence at the 3' end of the telomere. In 2009, ELIZABETN and others won the Nobel Prize in Medicine for their discovery of telomerase and the protective mechanism of telomeres on cell activity. Since then, many studies have shown that excessive activ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/48
CPCC12Q1/48C12Q2563/155
Inventor 何婧琳唐玲梅婷婷廖诗晴郭美彤
Owner CHANGSHA UNIVERSITY OF SCIENCE AND TECHNOLOGY
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