Research On Single Molecule Single Base Recognition Based On Solid Nanopores

In recent years,with the development of life science technology,DNA sequencing technology has attracted more and more attention,and has developed into a new discipline.After 20 years of development,the pore sensing detection technology as a new single molecule detection technology began to come into the mind of the researchers,is known as the third generation of DNA sequencing technology,different from the first two generation technology truly without tags,high flux of single molecule detection,it not only has high precision,also has a wide detection range and platform,including nucleic acid detection,identification and biological macromolecules combination of tumor markers detection and so on.Nanomaterials have received wide attention due to their unique physicochemical properties and good biocompatibility.Among them,gold nanoparticles,as a kind of nanomaterials with good biocompatibility,have also shown good application value in life science experiments.This paper mainly connect solid-state nanopores detection technology and nanoparticles assembly,implements the characteristic signal amplifier,explored in detail,small molecules,nucleic acid and protein detection of heavy metal ions from three aspects of the content by solid-state nanopores signal differences,for solid-state nanopores sensors in heavy metal ions detection and sensing test of markers associated with the disease to provide theoretical guidance.The main contents are as follows:1.Solid-state nanopores have long been a hotspot in the field of single molecule detection due to their stable and efficient properties.Due to the limitations of the manufacturing process,people have been trying to reduce their pore size to the size of biological nanopores but have not found a universal Methods.Here,we have developed a new SNP detection method based on solid-state nanopores.We use solid-state nanopore sensing technology to achieve high specificity for SNP,simple and fast detection.Through the design of the two-stage probe,the detection object is combined with the gold nanosphere,so that the nucleic acid signal that is not detected by the solid nanopore is converted into a large signal of the assembly,the signal amplification function is realized,and the solid nanopore detection platform is expanded.2.Mercury ions are closely related to People's Daily life and life health and safety.It is of great significance for environmental monitoring and people's life safety to develop effective detection methods of mercury ions and realize easy and rapid detection of mercury ions in the environment.In this work,we developed a new rapid mercury ion detection method based on solid-state nanopore biosensors.This experiment involves using the Au-DNA probe system and using the specific strong affinity of Hg²⁺and T bases to form a composite structure of T-Hg²⁺-T to capture mercury ions,successfully solving the problem that nanopores cannot directly detect mercury ion signals.3.We conducted a study on the blocking signal generated by the protein in the natural state and the protein in the denatured state when passing through the nanopore.At the same time,we screened and analyzed a large number of signals detected,and we found that the signal Blocking current and translocation time to determine the current structural state of the protein.The folded protein occupies a larger excluded volume in the nanopore,so it blocks more ion current than the protein in the unfolded or random coil state.Such findings can provide theoretical guidance for the pathogenesis of protein conformation diseases and provide technical support for the diagnosis and treatment of diseases.