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Phospholipid bilayer membrane for nanopore sequencing and preparation method thereof

A nanopore sequencing and double-layer membrane technology, applied in the field of biological sciences, can solve the problems of difficult repeated preparation, poor stability of phospholipid double-layer membrane, etc. Effect

Active Publication Date: 2021-09-17
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Aiming at the above-mentioned deficiencies in the prior art, the present invention provides a phospholipid bilayer membrane for nanopore sequencing and a preparation method thereof. Hydrophobic small molecule monomers are doped with phospholipids and polymerized in situ to solve the problems of poor stability of phospholipid bilayer membranes and difficulty in repeated preparation in the prior art

Method used

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  • Phospholipid bilayer membrane for nanopore sequencing and preparation method thereof
  • Phospholipid bilayer membrane for nanopore sequencing and preparation method thereof

Examples

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Embodiment 1

[0038] A phospholipid bilayer membrane for nanopore sequencing, the preparation method of which comprises the following steps in sequence:

[0039] S1: Take the polycarbonate pretreated by SU-8 as the nanoporous substrate, and fluorinate it with perfluorooctyltrichlorosilane to obtain a nanoporous substrate with stronger hydrophobicity;

[0040] S2: The free radical inhibitor in EHMA and TEGDMA is removed by alumina column, then, it and photoinitiator 2,2-diethoxy-1-phenylhexanone (DEAP) press 1:1:1 mole Mix the mixture to obtain a monomer mixture, then mix the monomer mixture with an equimolar amount of diphytylphosphatidylcholine (DPhPC) amphoteric phospholipid molecules and dissolve them in physiological saline to obtain a monomer-doped amphoteric phospholipid molecule solution.

[0041] S3: apply the monomer-doped amphoteric phospholipid molecular solution onto the modified nanopore, and dry with nitrogen gas;

[0042] S4: Add 1 mL of buffer solution on both sides of the ...

Embodiment 2

[0047] The difference between Example 2 and Example 1 is that the molar ratio of DPhPC:EHMA:TEGDMA:DEAP is 1:0.5:0.5:0.5.

Embodiment 3

[0049] The difference between Example 3 and Example 1 is that the molar ratio of DPhPC:EHMA:TEGDMA:DEAP is 1:2:2:2.

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Abstract

The invention discloses a phospholipid bilayer membrane for nanopore sequencing and a preparation method thereof, and the preparation method sequentially comprises the following steps: (1) mixing a hydrophobic micromolecule monomer, a photoinitiator and phospholipid molecules to obtain a phospholipid mixture; (2) smearing the phospholipid mixture on the nanopore base material, and volatilizing the solvent to obtain a phospholipid membrane; and (3) adding an ion buffer solution into bathing pools on two sides of the nanopore base material to redissolve phospholipid molecules, then recoating the nanopore base material with the phospholipid mixture, and carrying out photo-initiation polymerization to obtain the phospholipid bilayer membrane for nanopore sequencing. The invention also comprises the phospholipid bilayer membrane for nanopore sequencing prepared by the method. According to the invention, the hydrophobic micromolecular monomer with double bonds and the phospholipid are doped and subjected to in-situ polymerization, so that the problems of poor stability, difficulty in repeated preparation and the like of the phospholipid bilayer membrane in the prior art are solved.

Description

technical field [0001] The invention relates to the technical field of biological sciences, in particular to a phospholipid bilayer membrane for nanopore sequencing and a preparation method thereof. Background technique [0002] DNA is the code and blueprint that make up life. The sequence of bases in nucleotides in DNA constitutes genetic information, which can form RNA through the process of transcription, and then the mRNA in it can produce polypeptides and proteins through translation. As a relatively stable modification state, DNA methylation can be inherited to newborn offspring DNA during the DNA replication process under the action of DNA methyltransferase, which is an important epigenetic mechanism; therefore, as The only methylated form in mammals, 5-methylcytosine is also known as the "fifth base" of the human body. Abnormal DNA methylation is closely related to the occurrence and development of various diseases. Therefore, accurate detection of base sequence an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J9/36C08J9/40C08L69/00
CPCC08J9/36C08J9/405C08J2369/00C08J2433/10
Inventor 王云兵李高参徐鸿马博轩杨立
Owner SICHUAN UNIV
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