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DNA liquid state test method based on atomic force microscope

A technology of atomic force microscope and detection method, which is applied to measurement devices, preparation of test samples, instruments, etc., to achieve the effects of easy control, good stability and simple preparation method.

Inactive Publication Date: 2009-12-30
SHANGHAI NAT ENG RES CENT FORNANOTECH
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Problems solved by technology

However, most of these methods are limited to the imaging of DNA molecules in the atmosphere, and many DNA-related life processes can only reflect the real situation in the organism when observed in solution, so the development of simple, feasible and reliable AFM for DNA molecules in solution Imaging methods are the basis for better use of atomic force microscopy to observe DNA-related life processes and understand DNA-related biological responses

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[0023] The first step, take by weighing 203.0mg analytically pure magnesium chloride hexahydrate (MgCl 2 .6H 2 0), dissolved in a small beaker at room temperature, then transferred to constant volume in a 100.0mL volumetric flask, to obtain a concentration of magnesium chloride (MgCl) that was 10.0mmol / L 2 ) solution for use, take 10 μL of the original pUC18 DNA sample (concentration: 0.5 μg / μL) and dilute it to a concentration of 5.0 ng / μL (1.0 mL) with 10.0 mmol / L magnesium chloride solution to make a DNA sample solution;

[0024] In the second step, take 20.0 μL of the DNA sample solution and drop it on the freshly peeled mica sheet, absorb it for 1 minute, then wash it with ultrapure water, and directly add a 12 mm diameter polystyrene filter to the wet mica sheet without blowing it dry with nitrogen. The tetrafluoroethylene ring was pressed on the mica sheet to form a solution pool, and then 400.0 μL of HEPES buffer solution was added dropwise into the solution pool.

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Abstract

A DNA liquid state test method based on an atomic force microscope in the field of measurement for biotechnique comprises the following steps: configurating a buffer solution, then diluting a DNA sample solution to be observed to prepare the DNA sample solution; dripping the DNA sample solution on a mica substrate, washing the mica substrate with ultrapure water after adsorbing, finally setting an imaging test pool directly on the mica substrate; adding an imaging buffer solution into the imaging test pool followed by directly using the atomic force microscope to carry out scanning imaging process in the imaging test pool. The invention can realize AFM imaging of DNA molecules in the buffer solution in the absence of bivalent ions, thus being capable of avoiding the observation of the biological reaction process related to the DNA needing avoiding the participation of bivalent ions in some buffer solutions and providing an effective means for testing the conformation of the DNA molecules in life process.

Description

technical field [0001] The invention relates to a detection method in the technical field of biological detection, in particular to a DNA liquid state detection method based on an atomic force microscope. Background technique [0002] In 1985, Binnig cooperated with C.F.Quate of Stanford University and Christoph Gerber of IBM Zurich Laboratory to launch the Atomic Force Microscopy (AFM for short). Resolving imaging, it can obtain the shape of matter on the nanometer scale, and at the same time detect the interaction force between molecules, and there is no requirement for the conductivity of the sample, so it is widely used in the field of life sciences. After the appearance of AFM, it has been widely used in the field of biology, and has attracted the attention and favor of biologists. It can observe the structural characteristics of biomacromolecules at the single-molecule level, and at the same time reveal the information of the interaction between individual biomolecule...

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Application Information

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IPC IPC(8): G01N13/16G01N1/38
Inventor 沈新程刘晓晟许建荣何丹农
Owner SHANGHAI NAT ENG RES CENT FORNANOTECH
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