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Single particle biological probe and construction method for plasma biological storage device thereof

A technology of biological probes and construction methods, which can be applied to instruments, scientific instruments, measuring devices, etc., can solve the problems of low content of microRNAs and large size of information storage devices, and achieves improved mass transfer rate, better structure and stability, and space The effect of strong positioning ability

Active Publication Date: 2018-07-17
NANJING UNIV OF POSTS & TELECOMM +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, due to the low content of microRNAs, the detection limit of miR-21 can only reach 1fM for the plasmonic nanobiosensors constructed with probe molecules based on simple ssDNA-modified single metal nanoparticles. At the same time, the existing information storage devices The volume is relatively large, and there is still room for improvement

Method used

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  • Single particle biological probe and construction method for plasma biological storage device thereof
  • Single particle biological probe and construction method for plasma biological storage device thereof
  • Single particle biological probe and construction method for plasma biological storage device thereof

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Embodiment 1

[0063] This embodiment relates to a method for preparing tsDNA, comprising the following steps:

[0064] The design of tsDNA is based on the principles of complementary base pairing, as few secondary structures as possible and GC content above 50%, and three different sizes of tsDNA with side lengths of 7bp, 17bp and 26bp were designed. For the synthesis of tsDNA, the preparation method can refer to "A DNA nanostructure-based biomolecular probe carrier platform for electrochemical biosensing, Pei H, Lu N, Wen Y, et al. Advanced Materials, 2010, 22(42):4754-4758", the specific method as follows:

[0065] 1. Dissolve each strip of ssDNA, measure the absorption at 220-280nm under an ultraviolet spectrophotometer, obtain the molar extinction coefficient of each strip of ssDNA from Dalian Bao Biological Company, and determine the actual concentration of each strip of ssDNA;

[0066] 2. Mix the four ssDNAs forming tsDNA in equal proportions in TM buffer, add tris(2-carboxyethyl)pho...

Embodiment 2

[0074] This embodiment relates to a method for preparing an indium tin oxide conductive film glass substrate with Au@AgNCs fixed on the surface, comprising the following steps:

[0075] 1. Synthesis of nano-gold seeds: We used a common seed growth method to prepare gold seeds with a diameter of 20-40nm. In order to prepare gold nanoparticles with a diameter of 1-5 nm, first add 0.2-0.8 mL of fresh 5-15 mM sodium borohydride solution prepared in ice water to 5-15 mL of 50-150 mM CTAB and 0.2-0.5 mL of 5- In the mixed solution of 10mM chloroauric acid, the brown solution obtained was kept in a water bath at 20-30°C for 2-5 hours to ensure that the excess sodium borohydride was decomposed completely before use. Then, gold seed solution is made by following method, the 50~200mM ascorbic acid solution of 2~5mL is added in the 100~200mM CTAC solution of 5~15mL and the chloroauric acid mixed solution of 5~10mL 0.2~0.5mM to obtain After stirring evenly, add 0.2-0.5mL of 1-5nm gold na...

Embodiment 3

[0081] This embodiment relates to a method for modifying Au@Ag NCs by tsDNA, comprising the following steps:

[0082] Take 100-200 μL of 1pM tsDNA solution synthesized in Example 1 and drop it on the surface of the indium tin oxide conductive film glass with Au@Ag NCs immobilized in Example 2, and incubate at room temperature for 2-6 hours. Then the excess tsDNA solution was removed with ultrapure water and dried with nitrogen gas to prepare a single Au@Ag NC LSPR probe based on tsDNA.

[0083] Named according to the size of the side length of the tetrahedron, three tetrahedrons with different side lengths of 7bp, 17bp and 26bp are tsDNA 7 , tsDNA 17 and tsDNA 26 . Due to the strong covalent interaction between silver and sulfur, tsDNA with a side length of 17bp 17 For example, such as Figure 7 shown in tsDNA 17 thiol groups are modified on the three vertices of , one of the edges (l 1 ) to leave a section of ssDNA molecules for capturing miR-21 molecules, and on the o...

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Abstract

The invention provides a single particle biological probe and a construction method for a plasma biological storage device thereof. The construction method comprises the following steps: preparing a gold and silver core-shell nanometer cube (Au@AgNC2) sol and a tsDNA solution respectively, and fixing the Au@AgNC2 to the surface of indium tin oxide conducting film glass; dropwise adding the tsDNA solution to the surface of the indium tin oxide conducting film glass fixed with the Au@AgNCs, after incubation at room temperature, washing with ultrapure water, drying with nitgrogen, and thus obtaining the single particle LSPR biological probe based on tsDNA. Compared with a nanometer cube, the Au@AgNCs has similar plasma characteristics and more stable structure; and compared with a single-chain DNA molecule of a one-dimensional structure and a hairpin type DNA molecule of a two-dimensional structure, the tsDNA of a three-dimensional structure has the advantages of higher rigidity, structure stability, easy multi-functionalization and the like.

Description

technical field [0001] The invention relates to a construction method and application of a single-particle LSPR (localized surface plasmon resonance) probe based on tsDNA (tetrahedral structure DNA), belonging to the technical field of biological detection. Background technique [0002] MicroRNAs are a class of endogenous non-protein-coding RNAs with a length of 20-24 nucleotides, which usually play a very important role in early development, cell proliferation, differentiation, apoptosis and other biological processes. Abnormal expression of MicroRNAs occurs in precancerous malignant tumor cells and is associated with many cancers (such as: lung, liver cells, large intestine and breast cancer, etc.). MicroRNA-21 (miR-21) exists in many human cancer tissues mentioned above, especially the expression level in lung squamous cell carcinoma tissues is twice as high as that in normal tissues. Therefore, microRNAs can be used as biomarkers for the diagnosis and prognosis of early...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/25G01N21/47
CPCG01N21/25G01N21/47G01N2021/258
Inventor 汪联辉张颖帅振华翁丽星张磊周浩
Owner NANJING UNIV OF POSTS & TELECOMM
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