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A method for simultaneously distinguishing single-base mismatches at the single-molecule level

A base mismatch, single-molecule technology, applied in the direction of material electrochemical variables, can solve the problems of complex operation, high false positives, inability to achieve simultaneous distinction, etc., and achieve the effect of simple operation, low cost, and high thermodynamic stability.

Active Publication Date: 2020-11-10
SICHUAN UNIV
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

[0004] Complicated operation, time-consuming and expensive instruments, high false positive, especially only unique characteristic signal, unable to achieve simultaneous distinction

Method used

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  • A method for simultaneously distinguishing single-base mismatches at the single-molecule level
  • A method for simultaneously distinguishing single-base mismatches at the single-molecule level

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Embodiment Construction

[0019] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0020] The application principle of the present invention will be described in detail below in conjunction with the accompanying drawings.

[0021] like figure 1 As shown, the method for simultaneously distinguishing single-base mismatches at the single-molecule level provided by the embodiments of the present invention includes the following steps:

[0022] S101: Combined with nanopore electrochemical technology, a non-labeled co-arm modified molecular beacon is designed;

[0023] S102: Based on the characteristic blocking time of the perforation behavior of a single target molecule caused by molecular beacons, develop a...

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Abstract

The invention belongs to the field of recognition of single base mismatches of nucleic acid molecules, and discloses a method for simultaneously distinguishing single base mismatches at the single molecule level. The method for simultaneously distinguishing single base mismatches at the single-molecule level includes: combining nanopore electrochemical technology to determine a non-labeled co-arm-type improved molecular beacon; Single-molecule strategies simultaneously discriminate between perfectly complementary and single-base mismatched sequences. The present invention combines nanopore electrochemical technology to design a non-marked co-arm type improved molecular beacon, which shows a higher signal-to-background ratio (~104), higher thermodynamic stability and faster hybridization rate. This label-free single-molecule strategy realizes the simultaneous distinction of fully complementary and single-base mismatched sequences at the single-molecule level, and the invention has potential application value in the fields of rapid detection, accurate identification, and early clinical diagnosis.

Description

technical field [0001] The invention belongs to the field of recognition of nucleic acid base mismatches, in particular to a method for simultaneously distinguishing single base mismatches at the single molecule level. Background technique [0002] At present, the existing technologies commonly used in the industry are as follows: single base mutations are closely related to human diseases, and the identification of single base mismatches is of great significance for early disease diagnosis, pharmacogenetics and population genetics. Sequencing is a powerful technique sufficient to identify single base mismatches, but requires time costs (2-3 weeks), expensive instruments, and skilled laboratory skills. The polymerase chain reaction (PCR) proposed in recent years is a very sensitive single base mismatch detection technology, but it requires enzyme reaction and amplification of the target, which increases the complexity and cost of the experiment, which limits its practicality...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/26
CPCG01N27/26
Inventor 周翠松杨秋芳
Owner SICHUAN UNIV
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