DNA probe combined with gold nanospheres and application of DNA probe to detection of Hg<2+> based on solid-state nanopores

A technology of DNA probes and gold nanospheres, applied in the field of DNA probes combined with gold nanospheres, can solve the problems of high cost and cumbersome Hg2+ detection process, and achieve high sensitivity, convenient probe modification, and low-cost effects

Pending Publication Date: 2020-10-16
NANJING UNIV OF POSTS & TELECOMM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Purpose of the invention: the technical problem to be solved in the present invention is to provide a method for detecting

Method used

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  • DNA probe combined with gold nanospheres and application of DNA probe to detection of Hg&lt;2+&gt; based on solid-state nanopores
  • DNA probe combined with gold nanospheres and application of DNA probe to detection of Hg&lt;2+&gt; based on solid-state nanopores
  • DNA probe combined with gold nanospheres and application of DNA probe to detection of Hg&lt;2+&gt; based on solid-state nanopores

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Embodiment 1

[0033] In the present embodiment, the method for making a gold nanosphere carrier for modifying a DNA probe comprises the following steps:

[0034] Step 1: Design and prepare DNA probe sequences as follows:

[0035] (5'aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaactgcccattattattatta3'). One end of the binding chain is a polyA base polyA sequence, which is convenient for adsorption and wrapping with gold nanoparticles. The other end of the recognition chain is a sequence rich in T bases, which is easy to interact with Hg 2+ Form T-Hg 2+ -T compound structure, and at the same time, a small amount of A bases are doped in the recognition chain sequence, and the A bases can effectively avoid the self-hybridization of the probe.

[0036] Step 2: Take an appropriate amount of DNA probe solution and gold nanospheres with a particle size of about 4-6nm, mix them in a certain proportion, shake them well, and incubate them overnight in an incubator at 23-27°C at a speed of 200-400rpm.

[...

Embodiment 2

[0043] Add Hg in this embodiment 2+ Before and after, different gold nanosphere solutions are detected in the nanopore sensor, the specific method is as follows:

[0044] Step 1: Take an appropriate amount of the probe sample prepared above and add different concentrations of Hg 2+ After fully shaking the solution, incubate in an incubator at 23-27°C, 200-400rpm for different periods of time. The cultured samples are centrifuged and observed with a transmission electron microscope. It can be seen that the solution containing Hg 2+ Before and after the solution, the number of monomers and dimers changed greatly, proving that the designed probe has a strong effect on Hg 2+ Has a good capture effect.

[0045] Step 2: Translocate the assembled gold nanosphere solution in the nanopore. According to the size of the gold nanospheres, a nanopore sensor with a pore size of 40-60 nm is used to detect the assembly product. A solution is injected into one side of the fluidic device, a...

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Abstract

The invention discloses a probe using gold nanospheres as a carrier and application of the probe to detection of Hg<2+> based on solid-state nanopores, and belongs to the technical field of heavy metal detection. The gold-nanosphere-combined probe is used to identify Hg<2+> through a solid-state nanopore sensor, so that the highly sensitive detection of mercury ions by the solid-state nanopore sensor is realized. The difficulty that ions are directly to detect due to pores are too large in solid nanopore signal recognition is overcome, by taking gold nanoparticles as a carrier to carry out signal amplification, the high sensitive detection of the solid-state nanopores on Hg<2+> is realized, and the application of the solid-state nanopores in a molecular detection field is expanded.

Description

technical field [0001] The invention belongs to the technical field of heavy metal detection, in particular to a DNA probe combined with gold nanospheres and its detection of Hg based on solid-state nanopores. 2+ in the application. Background technique [0002] With the intensification of human industrialization, the content of heavy metals in the environment is increasing, and the pollution is getting worse. Among them, mercury ions are particularly serious in environmental pollution. It is one of the most common and most toxic heavy metals. It not only endangers the environment, but also can pass through The impact of the food chain poses a threat to human life. Metallic mercury mainly enters the respiratory tract of the human body in the form of vapor, and mercury compounds can enter the human body through the digestive tract, respiratory tract and skin. It can be seen that mercury ions can easily affect our life, health and safety, so the development of effective merc...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/6825
CPCC12Q1/6825C12Q2563/137C12Q2563/155C12Q2565/631C12Q2565/607C12Q2525/173
Inventor 张磊席国豪武灵芝叶媛
Owner NANJING UNIV OF POSTS & TELECOMM
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