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Method for detecting polymorphism of mononucleotide by using biotin-avidin system

A technology of single nucleotide polymorphism and biotin, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problem of no discovery, and achieve the effect of low cost

Inactive Publication Date: 2011-02-16
SHANGHAI JIAO TONG UNIV +1
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Problems solved by technology

[0006] In the further search of the prior art literature, there was no report on the detection of single nucleotide polymorphisms using the technology of "biotin-avidin system constructed on asymmetric two-dimensional DNA origami chip"

Method used

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Embodiment

[0029] Detection conditions:

[0030] In this embodiment, the shape of the map of China constructed by DNA origami is formed by repeated folding of the scaffolding long chain (single-stranded DNA of M13mp18 bacteriophage, up to 7249 bases), and the pattern of the map comes from the A set of 229 short staple chains. The feature of this graph is that 229 staple chains are more than two hundred addressable potential reaction sites, and any point on the template can be precisely positioned using the graph itself as a reference without index marks. In addition, any one of the 229 short DNA single strands can extend out the nucleotide sequence as a capture probe to hybridize with the complementary nucleotide sequence labeled with the nanoparticle, and serve as the labeling point of the nanoparticle on the origami pattern. Under the atomic force microscope (Atomic Force Microscope, AFM), there are prominent bright spots for observation.

[0031] In this example, the map of China wa...

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Abstract

The invention provides a method for detecting polymorphism of mononucleotide by using a biotin-avidin system in the technical field of nanometer. The method comprises the following steps: constructing an asymmetric two-dimensional graph by utilizing a DNA paper folding art to prepare a DNA paper folding chip; hybridizing a report probe of a tail end modified biotin with the DNA paper folding chip; connecting streptavidin with the biotin; detecting the polymorphism of the mononucleotide; and imaging by an atomic force microscope. The method has the advantages of no need of restrict experiment conditions or complicated enzyme reaction, simpleness, low cost and the like. By using the method, the DNA hybridizing process can be realized in nanometer scale and liquid phase reaction, and high sensitivity biomedical detection can be reached.

Description

technical field [0001] The present invention relates to a detection method in the technical field of bioengineering, in particular to the detection of single nucleosides using a biotin-avidin system (Biotin-Avidin-System, BAS) constructed on an asymmetric two-dimensional DNA origami chip. Acid polymorphism method. Background technique [0002] The biotin-avidin system is a new biological reaction amplification system developed in the late 1970s. With the advent of various biotin derivatives, BAS was soon widely used in various fields of medicine. A large number of studies in recent years have confirmed that the biotin-avidin system can be combined with almost all kinds of markers that have been successfully studied so far. The strong combination of biotin-avidin and the high affinity of the labeling reagent and the multi-stage amplification effect make BAS immunolabeling and related tracer analysis more sensitive. It has become a new technology that is widely used in the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 贺林李璨张钊陈培华
Owner SHANGHAI JIAO TONG UNIV
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