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Accurate single-molecule force spectroscopy method with high throughput

A single-molecule force spectroscopy, high-throughput technology, applied in the field of precise high-throughput single-molecule force spectroscopy, can solve the problems of low detection efficiency, poor specificity and accuracy, and achieve high detection efficiency, specificity and accuracy Enhanced, efficient and precise measurement of the effect

Active Publication Date: 2019-04-02
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 an accurate 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

Method used

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  • Accurate single-molecule force spectroscopy method with high throughput
  • Accurate single-molecule force spectroscopy method with high throughput
  • Accurate single-molecule force spectroscopy method with high throughput

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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 an accurate single-molecule force spectroscopy method with high throughput. The method comprises the following steps of: 1) using DNA origami as a substrate, selecting several modification sites on the DNA origami according to the geometry of the DNA origami, and extending a stretch of a target DNA strand at the 5' end of the oligonucleotide strand of the selected modification site, wherein the target DNA strand can be complementary to the DNA strand immobilized on an AFM probe; and 2) measuring the interaction force between two complementary DNA strands located on the DNA origami and the AFM probe by an atomic force microscopy single-molecule force spectroscopy method, and collecting the specific force-distance curve data. According to the accurate single-molecule force spectroscopy method with high throughput, the detection efficiency is high, and the specificity and precision are improved. The simple and efficient single-molecule force spectroscopy method withhigh throughput provides an efficient and accurate measuring method for the quantitative measurement of single molecule mechanics.

Description

technical field [0001] The invention relates to the field of single-molecule force spectrum measurement, and more specifically relates to a precise 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 a...

Claims

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Application Information

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Patent Type & Authority Applications(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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