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Preparation method of pH sensitive element based on DNA molecule configuration change

A DNA molecule and configuration change technology, applied in the field of DNA molecules, to achieve high research significance and application value, satisfying the effect of mature detection and material preparation technology

Inactive Publication Date: 2015-03-04
SHANGHAI NAT ENG RES CENT FORNANOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the three-dimensional origami structure of DNA is a kind of biocompatibility, stable structure, and response-sensitive bio-controlled release material, and its sensitive response to pH value has not been applied to detection or any other field.

Method used

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  • Preparation method of pH sensitive element based on DNA molecule configuration change
  • Preparation method of pH sensitive element based on DNA molecule configuration change
  • Preparation method of pH sensitive element based on DNA molecule configuration change

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Take 2 μL single-stranded DNA (50 μM) 1, 2, 3, 4 and add to 42 μL Tris-MgCl (Tris 10mM, MgCl 2 50mM, pH8) solution. The mixed solution was placed in a PCR instrument with a reaction temperature of 95° C., cooled to 4° C. after 10 minutes, and the reaction was continued for 30 minutes. A single strand of DNA can self-assemble into a DNA tetrahedron containing i-motif through complementary base pairing.

[0031] Take 100 μL of thiol coupling reagent (SH-(PEG) 7 -OCH 3 , 0.1mg / mL) was added dropwise on the surface of the gold-plated quartz crystal, and left to stand for 30 minutes. Take 100 μL of the self-assembled DNA molecule solution, drop it on the surface of the gold-plated quartz crystal, and fully modify it for 5 hours (overnight).

[0032] The TDN i-motif molecule is subjected to H under acidic conditions + The effect of DNA tetrahedron will be deformed to a certain extent. At the same time, the i-motif segment will be due to the H + The role of folding. T...

Embodiment 2

[0034] Take 2 μL single-stranded DNA (50 μM) 2, 3, 4, 5 and add to 42 μL Tris-MgCl (Tris 10mM, MgCl 2 50mM, pH8) solution. The mixed solution was placed in a PCR instrument with a reaction temperature of 95° C., cooled to 4° C. after 10 minutes, and the reaction was continued for 30 minutes. A single strand of DNA can self-assemble into a DNA tetrahedron through complementary base pairing.

[0035] Take 100 μL of thiol coupling reagent (SH-(PEG) 7 -OCH 3 , 0.1mg / mL) was added dropwise on the surface of the gold-plated quartz crystal, and left to stand for 30 minutes. Take 100 μL of the self-assembled DNA molecule solution, drop it on the surface of the gold-plated quartz crystal, and fully modify it for 5 hours (overnight).

[0036] The gold-coated quartz crystal TDN molecules are subjected to H under acidic conditions + The effect of DNA tetrahedron will be deformed to a certain extent. Therefore, the TDN molecule can intuitively reflect the change of pH on the change ...

Embodiment 3

[0038] Take 2 μL single-stranded DNA (50 μM) 6 and 7 and add to 46 μL Tris-MgCl (Tris 10 mM, MgCl 2 50mM, pH8) solution. Single-stranded DNA can self-assemble into double-stranded DNA molecules through complementary base pairing.

[0039] Take 100 μL of thiol coupling reagent (SH-(PEG) 7 -OCH 3 , 0.1mg / mL) was added dropwise on the surface of the gold-plated quartz crystal, and left to stand for 30 minutes. Take 100 μL of the self-assembled DNA molecule solution, drop it on the surface of the gold-plated quartz crystal, and fully modify it for 5 hours (overnight).

[0040] The gold-coated quartz crystal-double-stranded DNA molecule is subjected to H under acidic conditions + The role of the double-stranded DNA molecules will be curled to a certain extent. Therefore, double-stranded DNA molecules can intuitively reflect the change of pH in the change of configuration.

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Abstract

The invention relates to a preparation method of a pH sensitive element based on DNA molecule configuration change. The preparation method comprises the following steps: preparing a DNA self-assembling structure; and adsorbing a DNA molecule and a gold base to obtain the pH sensitive element. The DNA molecule is subjected to the action of H<+1> under an acidic condition and a chain segment has the configuration change to a certain extent; and meanwhile, some DNA chain segments which are sensitive to pH, such as i-motig chain segments, are folded under the action the H<+1>, so that the change of the pH can be directly reflected through the change of the configuration by the DNA molecule. According to the preparation method, the DNA molecule is modified on a gold surface, a material preparation process is developed and the maneuverability is strong; the sensitivity to the change of the pH is high and a rapid reaction can be realized; and a proton-driven environment-friendly and clean reversible biological change-over switch device is provided and has a wide application prospect in the field of biosensors and the field of clean energy sources.

Description

technical field [0001] The invention proposes a preparation method of a pH sensitive element based on the configuration change of DNA molecules, and specifically relates to a DNA molecule with different configurations modified on the surface of gold as a substrate. Background technique [0002] The pH value plays a very important role in the normal operation of organisms, involving many physiological and pathological processes, such as cell proliferation and apoptosis, endocytosis, ion transport and dynamic balance, multidrug resistance, etc. Once the pH value Abnormal, will cause cell dysfunction. The pH distribution of the human body is extremely complex. The blood pH ranges from 7.35 to 7.45, the pancreas ranges from 8.0 to 8.3, and the gastric juice ranges from 0.8 to 1.5; the pH in cells can also be divided into two ranges, acidic organelles (endosomes, The pH range of lysosomes is 4.5~6.0, and that of cytoplasm is 6.8~7.4; in addition, the secretion of some tumor cell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 何丹农王萍夏智伟
Owner SHANGHAI NAT ENG RES CENT FORNANOTECH
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