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A high-throughput single-molecule force spectroscopy method

A single-molecule force spectroscopy and high-throughput technology, which is applied in measuring devices, instruments, scanning probe microscopy, etc., can solve the problems of low detection efficiency, poor specificity and accuracy, and achieve high detection efficiency and specificity. Improved performance and accuracy, simplified data analysis

Active Publication Date: 2021-05-18
SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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Problems solved by technology

[0005] The purpose of the present invention is to provide a high-throughput single-molecule force spectroscopy method, thereby solving the problems of low detection efficiency, poor specificity and accuracy in the single-molecule force spectroscopy method in the prior art

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  • A high-throughput single-molecule force spectroscopy method
  • A high-throughput single-molecule force spectroscopy method
  • A high-throughput single-molecule force spectroscopy method

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Embodiment 1

[0039] The DNA origami used in this example is a triangular DNA origami, which can be self-assembled into a triangular pattern with an external side length of 120 nm and an internal side length of 40 nm by using M13mp18 DNA and staple strands and a single strand of target DNA staple. That is, a DNA origami modified with single-strand DNA staples of interest. On the three sides A, B, and C of the triangle DNA origami, six modification sites were respectively selected, specifically as figure 2 As shown, a target DNA short chain is extended at the 5' end of the oligonucleotide chain at the selected modification site. The target DNA short chain can be 20 bases, which can be as short as the DNA immobilized on the AFM needle tip. The strands are complementary and paired, and then the interaction force between the two short DNA strands is measured by single-molecule force spectroscopy using an atomic force microscope.

[0040] 1.1 Preparation of triangular DNA origami with target D...

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Abstract

The invention provides a high-throughput single-molecule force spectroscopy method, which includes the following steps: 1) using DNA origami as a base, selecting several modification sites on the DNA origami according to the geometric structure of the DNA origami, and The 5' end of the spotted oligonucleotide chain extends a short target DNA strand, which can be complementary to the DNA short strand immobilized on the AFM probe; Methods The interaction force between two complementary DNA short strands located on DNA origami and AFM probes was measured, and the data of specific force-distance curves were collected. The present invention provides a high-throughput single-molecule force spectrum method with high detection efficiency and improved specificity and accuracy. This simple and efficient high-throughput single-molecule force spectrum method provides quantitative measurement of single-molecule mechanics. An efficient and accurate measurement method.

Description

technical field [0001] The invention relates to the field of single-molecule force spectrum measurement, in particular to a high-throughput single-molecule force spectrum method. Background technique [0002] For the interactions within and between biomolecules, such as changes in the internal structure of biomolecules and mutual recognition between molecules, mechanical properties can be used to qualitatively and quantitatively study. Detection methods at the single-molecule level are an inevitable requirement for the study of the above-mentioned forces. In the past few decades, with the rapid development of science and technology around the world, a variety of single-molecule force spectroscopy techniques have emerged, such as optical tweezers, magnetic tweezers, and single-molecule force spectroscopy based on atomic force microscopy. Wait. However, the main limitations of optical tweezers are: during measurement, the light beam will cause light and thermal damage to bio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01Q60/24
CPCG01Q60/24
Inventor 李宾刘文静张萍
Owner SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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