Non-labeled aptamer probe system and detection method and application thereof

A detection method and aptamer technology, applied in measuring devices, instruments, fluorescence/phosphorescence, etc., can solve problems such as signal leakage, achieve low detection cost, fast detection time, and reduce background signal interference

Active Publication Date: 2020-11-03
深圳立专信息科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the inventors have found that while the double strands are being produced during the reaction, there is also a problem of signal leakage

Method used

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  • Non-labeled aptamer probe system and detection method and application thereof
  • Non-labeled aptamer probe system and detection method and application thereof
  • Non-labeled aptamer probe system and detection method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment

[0082] 1. Test materials and methods

[0083] 1.1 Reagents

[0084] Table 1 Main materials and reagents

[0085]

[0086] Table 2 Nucleotide sequence list

[0087]

[0088]

[0089]Note: The part in italics in the table is the complementary region between CAP aptamer and Primer.

[0090] (1) PBS buffer solution (10μM)

[0091] Liquid A: 4.38g solid sodium chloride, 1.27g solid magnesium chloride, 7.8g solid sodium dihydrogen phosphate, 500mL. Liquid B: 4.38g sodium chloride solid, 17.91g disodium hydrogen phosphate solid, 500mL, store at 4°C for later use, mix liquid A and liquid B until the pH value is 7.4.

[0092] (2) 10×TE buffer solution

[0093] Weigh 1.21 g of Tris-Base solid and 0.372 g of EDTA solid with a balance, adjust the pH to 7.8, set the volume to 10 mL, and place at 4°C for use.

[0094] (3) CAP stock solution (100mg / L)

[0095] Weigh 0.01g of CAP solid with a balance, dilute to 100mL with buffer solution, and store in a refrigerator at 4°C for...

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Abstract

The invention provides a non-labeled aptamer probe system as well as a detection method and application thereof, and belongs to the technical field of biological detection and analysis. According to the present invention, the SYBR Green I fluorescent dye is adopted to dye the complementary base, and the design of the non-labeled aptamer probe system is performed, such that the signal amplificationis achieved, and the cost is low; through the design of the initiation chain, four hairpins can be initiated to form a large number of four-arm connectors so as to generate more complementary pairingbases, the four-arm connectors can be combined with more SYBR Green I, and cyclic amplification of signals is achieved while signal leakage is reduced; meanwhile, based on the characteristics of T7 exonuclease, hairpin enzymolysis which is not caused in an experiment is achieved, background signal interference is reduced, the sensitivity of CAP detection is improved, and good practical application value is achieved.

Description

technical field [0001] The invention belongs to the technical field of biological detection and analysis, and in particular relates to a non-marked aptamer probe system and a detection method and application thereof. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Chloramphenicol (CAP) is a neutral nitrobenzene derivative obtained from Streptomyces venezuel. It is usually used as a veterinary medicine to treat and prevent infections caused by various germs. However, while CAP brings benefits, there are also many hidden dangers, such as the generation of aplastic anemia, leukemia and gray baby syndrome. Therefore, based on the consideration of people's health, many count...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N27/447
CPCG01N21/6428G01N27/447
Inventor 贾敏唐琳刘雨婕陈爽
Owner 深圳立专信息科技有限公司
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