Nano-structure-based protein unimolecular electronic device and preparation and application thereof
A technology for electronic devices and proteins, which is used in biochemical equipment and methods, and the determination/inspection of microorganisms. Single-molecule sequencing, improving capture efficiency, and achieving the effect of continuous sequencing
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Embodiment 1
[0050] Embodiment 1: DNA polymerase molecular electronic device based on nanopore-counter electrode structure
[0051] (1) Preparation of nanopore-counter electrode structure: use a series of micro-nano processing methods such as photolithography, dry etching, wet etching, reactive ion beam etching, etc. 2 / SiN substrate is processed to prepare a suspended membrane structure 1 with a SiN film with a size of 2 square microns and a thickness of 30 nanometers, such as figure 1 shown. Among them, SiO 2 The film 1b acts as an insulating buffer layer to support the SiN film 1c and reduce the capacitive effect in electrical measurement; the Si substrate 1a mainly supports the SiO 2 The role of membrane 1b. The material of the suspended membrane structure 1 is not limited to Si / SiO 2 / SiN, and other materials such as graphene can also be used to prepare a suspended membrane structure. On the suspended film structure 1, the nano-counter electrode 3 is prepared by a conventional el...
Embodiment 2
[0058] Embodiment 2: RNA polymerase molecular electronic device based on nanopore-counter electrode structure
[0059] (1) Preparation of nanopore-counter electrode structure: Same as Example 1.
[0060] (2) The modification of the surface of the nano-pair electrode: the same as in Example 1.
[0061] (3) Preparation of nanopore-counter electrode-protein composite structure by chemical crosslinking method: inject 500 microliters of 1:1 mixed solution of NHS solution (100mM) and EDC solution (100mM) into the centrifuge tube, put For the chemically modified nanopore-counter electrode chip, 200 microliters of RNA polymerase (50 micrograms / ml) was dropped into a centrifuge tube, and stored in the dark for 1 hour, so that the RNA polymerase and the nanometer counter electrode were cross-linked. After the end, take it out, soak it in deionized water, and save it for later use. The nanopore-counter electrode-protein composite structure has the function of detecting DNA sequence.
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Embodiment 3
[0066] Embodiment 3: DNA exonuclease molecular electronic device based on nanopore-counter electrode structure
[0067] (1) Preparation of nanopore-counter electrode structure: Same as Example 1.
[0068] (2) The modification of the surface of the nano-pair electrode: the same as in Example 1.
[0069] (3) Preparation of nanopore-counter electrode-protein composite structure by chemical crosslinking method: inject 500 microliters of 1:1 (volume ratio) mixed solution of NHS solution (100mM) and EDC solution (100mM) into the centrifuge tube, put into the chemically modified nanopore-counter electrode chip, drop 200 microliters of DNA exonuclease (50 micrograms / ml) into a centrifuge tube, and store it in the dark for 1 hour, so that the DNA exonuclease and the chemically modified nano Gold cross-links the electrodes. After the end, take it out, soak it in deionized water, and save it for later use. The nanopore-counter electrode-protein composite structure has the function of ...
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