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A glass nanopore with a diameter less than 10nm, its preparation method and its application for detecting dna

A nanopore and glass technology, applied in the field of preparation and detection of DNA, in the field of glass nanopores, can solve the problems of irreversibility and high experimental cost, preventing practical use, inapplicability, etc., to improve the DNA transport signal and signal-to-noise ratio. , the effect of reducing the concentration, the method is simple

Active Publication Date: 2020-06-02
ANHUI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But it is not suitable for glass nanopores with aspect ratio ~104, because the tip length of glass nanopores is generally greater than 1 mm, which is much longer than these polymer nanopores.
The irreversibility of the preparation of the first method and the high experimental cost, while the second method prevents their widespread practical use due to the complexity of the method and its dependence on expensive vacuum equipment

Method used

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  • A glass nanopore with a diameter less than 10nm, its preparation method and its application for detecting dna
  • A glass nanopore with a diameter less than 10nm, its preparation method and its application for detecting dna
  • A glass nanopore with a diameter less than 10nm, its preparation method and its application for detecting dna

Examples

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Effect test

Embodiment 1

[0040] A method for preparing a glass nanopore with a diameter less than 10nm, comprising the following steps:

[0041] 1) Glass capillary inner diameter: 0.7mm; outer diameter: 1mm; length 10cm; purchased from Sutter Company; install the glass capillary on the P-2000 laser microelectrode puller, stabilize the two moving rods with aluminum fixtures, and pass the following Heating program: heating=550°C, filament=3, speed=99m / s, delay=20ms, pull=50N, heating program is applied for 40 seconds, then 20 seconds cooling period, the above-mentioned heating and cooling process is repeated 3-4 times to obtain Microporous preforms.

[0042] 2) Remove the fixture from the puller, and perform the following pulling program to pull the preform into two glass nanopores with a diameter less than 10 nm: Pulling program parameters: heating = 660 ° C, filament = 1, speed = 60m / s, delay = 165ms, pull = 225-230N, pull off, that is, two glass nanopores with a diameter of less than 10nm are obtain...

Embodiment 2

[0052] The application of a glass nanopore with a diameter less than 10nm prepared in Example 1 for detecting DNA, the specific method is:

[0053] Chemical experiments such as cyclic voltammetry (CV) and chronoamperometry (I-t) were performed on a CHI 660C (Shanghai Huachen) electrochemical workstation using a two-electrode system. By inserting the Ag / AgCl electrode into the prepared nanopore tail as the working electrode; the counter electrode is also an Ag / AgCl electrode, immersed in the solution (1.0M KCl in 10mM PBS, pH7.4) outside the nanopore. Please refer to the illustration for details figure 2 . Cyclic voltammetry curves CV and current-time (I-t) curves were amplified and collected by using a current amplifier (Axon 200b, Axon Instrument, Forest City, USA) and Shanghai Chenhua Electrochemical Workstation. The detected DNA is λ-DNA (pBR322-DNA) purchased from Shanghai Sangon Bioengineering Co., Ltd.

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Abstract

The invention provides a glass nanopore with a diameter less than 10nm, a preparation method and an application thereof for detecting DNA, directly optimizing a drawing parameter and a drawing processof a laser drawing device, and reducing the diameter of the conical glass capillary nanopore. By this method, glass micropore nanopores smaller than 10 nm in diameter can be easily obtained. The method is simple, direct and environmentally friendly, and does not rely on expensive vacuum equipment. The shrinkage of the glass capillary nanopore prepared by the invention not only reduces the concentration of detectable DNA, but also remarkably improves the DNA transporting signal and the signal-to-noise ratio, so that the method and the glass nanopore are expected to be used for DNA transportingresearch.

Description

technical field [0001] The invention belongs to the field of nanomaterial preparation, and in particular relates to a glass nanopore with a diameter less than 10 nm, a preparation method and an application for detecting DNA. Background technique [0002] With the rapid development of life sciences, the research on life phenomena has gone deep to the level of single molecule and single cell. For this reason, analytical means having detection sensitivity at the single-molecule level are required to adapt to this development. Among many single-molecule detection technologies, the nanopore single-molecule detection technology based on the Coulter counter principle is simple and accurate, and shows great application potential in fast and cheap DNA detection, and has attracted increasing attention. [0003] Although biological nanopores were first studied and applied to detect DNA, people also found its shortcomings during the research process. For example, due to the poor stabi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B81B1/00B82Y40/00C12Q1/6869
CPCB81B1/004B82Y40/00C12Q1/68C12Q2565/631C12Q2563/137C12Q2565/607
Inventor 李永新杨程汤浩然汪浩陈唯
Owner ANHUI NORMAL UNIV
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