A detection kit based on nucleic acid aptamer

A detection kit and nucleic acid aptamer technology, applied in the field of biosensors, can solve the problems of difficult design, limited scope of use, complexity, etc., and achieve the effects of improving detection sensitivity, low cost, and simple operation

Active Publication Date: 2020-06-05
GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Existing nucleic acid aptamer-based detection methods are generally more complicated, and it is difficult to detect different substances by changing the nucleic acid aptamer sequence, which makes its design more difficult and limits its scope of application

Method used

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  • A detection kit based on nucleic acid aptamer
  • A detection kit based on nucleic acid aptamer
  • A detection kit based on nucleic acid aptamer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1: A kind of ATP detection kit comprises the following components:

[0052] (1) Nucleic acid H1, DNA1, DNA2, DNA3, the sequences of which are as follows:

[0053] H1: 5'-CGAATCAAGAGT(a)-ACCTGGGGAGTAT(b)-CACCGTATC(c)-CTATGCCTGCAACGT(d)-GATACGGTG(c*)-TGCGGAGGAAGGT(e)-CTACAAATAAGC(f)-3'

[0054] DNA1: 5'-GATATC(g)-AGCGATTAAC(Ⅰ)-ACTCTTGATTCG(a*)-3'

[0055] DNA2: 5'-GCTTATTTGTAG(f*)-GTTACACCCATGT(Ⅱ)-TACTCT(h)-3'

[0056] DNA3: 5'-FAM-AGAGTA(h*)-TrAG-GATATC(g*)-BHQ1-3'

[0057] (2) ATP standard solution

[0058] (3) Reaction buffer: containing 10mM Tris-HCl, pH 7.5, containing 50mMNaCl, 35mM MgCl 2 .

[0059] Its usage is as follows:

[0060] 1) Use reaction buffer (10mM Tris-HCl, pH 7.5, containing 50mMNaCl, 35mM MgCl 2 ) dissolve H1, DNA1, DNA2, DNA3 respectively;

[0061] 2) Add different concentrations of ATP into 200nM H1 solution, mix well, and react at room temperature for 45 minutes;

[0062] 3) Add 200nM DNA1 and DNA2 solutions, mix thoroughly, a...

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Abstract

The invention discloses a detection kit based on an aptamer. The detection kit comprises stem-loop structure nucleic acid H1, DNA (deoxyribonucleic acid) 1, DNA2 and DNA3, the stem-loop structure nucleic acid comprises the aptamer divided into two sections, the aptamer serves as a molecular recognition element, specific binding of molecules to be detected is achieved, catalytic activity of DNA enzymes is activated, substrate chains are circularly cut, high fluorescent detection signals are generated, and high-sensitivity detection is achieved. Detection limit of the detection kit for ATP (adenosine triphosphate) is 0.2 nanometer, detected linear range is wide and is 1 nanometer to 10 micrometers, a detection system has excellent specificity, common analogues comprising CTP (cytidine triphosphate), GTP (guanosine triphosphate) and UTP (uridine triphosphate) do not interfere detection results, separation and purification processes are omitted in the detecting process, detection can be achieved by simple blending, the detection kit has the advantages of simplicity in operation, low cost, quick response and the like, and the detection kit can be used for rapidly detecting and analyzingthe content of the ATP in environments or food samples.

Description

technical field [0001] The invention belongs to the field of biosensors and relates to a nucleic acid aptamer-based detection kit. Background technique [0002] Nucleic acid aptamer (aptamer) is an oligonucleotide fragment that can specifically bind to proteins or other small molecular substances screened by the in vitro screening technology SELEX (Systematic Evolution of Ligands by Exponential Enrichment). As a new type of molecular recognition tool, nucleic acid aptamer not only has high affinity and specificity, but also has the advantages of easy synthesis, easy modification, and good stability compared with traditional antibodies. It has a good application in the field of protein and molecular detection. application potential. In recent years, combined with the molecular characteristics of nucleic acid aptamers, the detection of proteins has been converted into the detection of specifically recognized nucleic acid aptamers, which not only avoids the shortcomings of tra...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/25
CPCC12Q1/25
Inventor 陈俊华潘家峰周丹华于焕云
Owner GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
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