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Method for preparing specimen in use for researching DNA through microscope in atomic force

An atomic force microscope and sample technology, applied in biochemical equipment and methods, microbial measurement/inspection, surface/boundary effects, etc., can solve problems such as uncontrollability, destruction, and waste of DNA samples

Inactive Publication Date: 2005-11-16
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the uncontrollability of this treatment method in the process of washing with water, the success rate of the prepared DNA sample is very low (destruction of the DNA conformation adsorbed to the substrate and partial clustering of the DNA, etc.)
Furthermore, short-term washing cannot effectively remove the salt ions adsorbed on the DNA molecules, so that they precipitate on the surface and DNA molecules during the drying process to form salt crystals, which seriously interfere with DNA imaging.
In addition, washing with water also removes a large number of DNA molecules, so that the DNA molecules adsorbed to the surface of the substrate become very small, resulting in the waste of DNA samples.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] 12.5ng / μl λ~DNA with 12.5mM Mg 2+ The mixed solution was heated at 38°C for 30 minutes, and 20 μl of it was dropped on the 1.5cm 2 Adsorb the newly dissociated mica surface for 7-8 minutes (25°C), diffuse the mica in ultra-pure water for 30 minutes at 4°C, take it out and put it in absolute ethanol for 30 seconds, and finally place it at 25°C under nitrogen protection Dry for 2 hours. Atomic force microscope characterization This method prepared DNA compact monomolecular film.

Embodiment 2

[0014] The mixed solution of 2.5ng / μl plasmid DNA pBR 322 and 2.5mM magnesium acetate was heated at 38°C for 30 minutes, and 20μl was dropped on 1.5cm 2 Adsorb the newly dissociated mica surface for 7-8 minutes (25°C), diffuse the mica in ultrapure water for 30 minutes at 4°C, take it out and place it in absolute ethanol for 30 seconds, and finally place it at 25°C under nitrogen protection Dry for 2 hours. Atomic Force Microscopy Characterization This method prepares expanded plasmid DNA molecules.

Embodiment 3

[0016] 2.5ng / μl linear DNA pBR 322 / PstI mixed with 2.5mM magnesium acetate was heated at 38°C for 30 minutes, and 20μl was dropped on 1.5cm 2 Adsorb the newly dissociated mica surface for 7-8 minutes (25°C), diffuse the mica in ultra-pure water for 30 minutes at 4°C, take it out and put it in absolute ethanol for 30 seconds, and finally store it at 25°C under nitrogen protection Dry for 2 hours. Atomic Force Microscopy Characterization This method produces fully extended linear DNA molecules.

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PUM

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Abstract

A process for preparing the specimen used to research DNA by atomic force microscope includes proportionally mixing DNA with mixed solution of two-valence metallic ions, heating, dripping it on the surface of newly dissociated mica, adsorption, diffusing inorganic salt ions in ultrapure water, immersing in absolute alcohol, and drying in N2.

Description

technical field [0001] The invention belongs to a method for preparing a sample by using a diffusion method in ultrapure water, in particular to a method for preparing a sample used in the study of DNA by an atomic force microscope. Background technique [0002] Atomic force microscope is one of the third generation of new microscopes developed after optical microscope and electron microscope. It relies on tiny probes to detect the shape of the measured object and its properties such as electricity, magnetism, viscoelasticity, and hardness. Due to its ultra-high resolution, the need for no conductive substrate, and the ability to operate in liquid and atmospheric environments, atomic force microscopy has been widely used to image DNA and study the interaction of DNA with other molecules. In these studies, mica has become a widely used substrate due to its atomic-level flatness, super-large single-crystal facets, and easy dissociation to prepare new surfaces. Therefore, the...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01Q30/20
Inventor 宋永海李壮刘志国魏刚王丽孙兰兰
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI