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Method for detecting ATP aptamer and ATP

A technology of aptamers and biochips, which is applied in biochemical equipment and methods, microbial measurement/testing, fluorescence/phosphorescence, etc., can solve the problems of only millimolar sensitivity, large sample consumption, and difficulty in miniaturization , to achieve the effect of wide detection linear range, short time consumption and simple method

Inactive Publication Date: 2014-04-16
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide a method for detecting ATP aptamers and ATP in order to solve the shortcomings of the existing methods for detecting ATP, such as complex operation, large sample consumption, difficulty in miniaturization and millimolar sensitivity.

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  • Method for detecting ATP aptamer and ATP
  • Method for detecting ATP aptamer and ATP

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example 1

[0033] 1. Mix the molar concentration ratio of ATP aptamer and poly-T chain at 1:3 with sample solution to obtain ATP aptamer solution, the concentration of ATP aptamer is 0.1 μmol / L, 0.5 μmol / L, 1 μmol / L L, 5 μmol / L, 10 μmol / L, 20 μmol / L, take 15 μL of the sample solution on the sample plate, and apply it on the SmartArrayer 48 biochip spotting system (the production method of the biochip application standard, select Beijing Boao Crystal core produced by Biotechnology Co., Ltd. Optical-grade aldehyde-based substrate), in order to obtain a good array point and maintain the activity of biomolecules, the composition of the sample spotting solution is: sodium dodecyl sulfate with a mass fraction of 0.005%, pH=7.0 containing 0.45mol / L sodium chloride, 45mmol / L trisodium citrate (dihydrate) buffer solution (PH=7.0) of 1.5mol / L betaine, it is 30 ℃ that the chip that spot sample is put into, and relative humidity is 80% In a constant temperature and humidity chamber, react for 12 ...

example 2

[0038] 1. Mix the molar concentration ratio of ATP aptamer and poly-T chain at 1:3 with sample solution to obtain ATP aptamer solution, the concentration of ATP aptamer is 0.1 μmol / L, 0.5 μmol / L, 1 μmol / L L, 5 μmol / L, 10 μmol / L, 20 μmol / L, take 15 μL of the sample solution on the sample plate, and apply it on the SmartArrayer 48 biochip spotting system (the production method of the biochip application standard, select Beijing Boao Crystal core produced by Biotechnology Co., Ltd. Optical-grade aldehyde-based substrate), in order to obtain a good array point and maintain the activity of biomolecules, the composition of the sample spotting solution is: sodium dodecyl sulfate with a mass fraction of 0.005%, pH=7.0 containing 0.45mol / L sodium chloride, 45mmol / L trisodium citrate (dihydrate) buffer solution (PH=7.0) of 1.5mol / L betaine, it is 30 ℃ that the chip that spot sample is put into, and relative humidity is 80% In a constant temperature and humidity chamber, react for 12 ...

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Abstract

A method for detecting an ATP aptamer and ATP belongs to the technical field of biology and overcomes the defects that the conventional ATP detecting method is complicated in operation, large in sample consumption and difficult to miniaturize and only has a sensitivity of millimolar level. Through a DNA microarray technology, the ATP aptamer is fixed, a DNA complementary strand with a fluorescent dye CY5 is taken as a marker, and a biochip is marked through a hybridization reaction; through specific recognition of the ATP and the ATP aptamer and in combination with a microarray fluorescent scanner, a fluorescent signal of the fluorescent dye on the biochip can be obtained and the synchronous detection of the ATP aptamer and the ATP can be realized. The method is simple; the sensitivity can reach 1 nM; the ATP aptamer and the ATP can be detected at the same time; the detection limit of the ATP is 1nM; and the linear range is 3 orders of magnitude.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for detecting ATP aptamers and ATP. Background technique [0002] Adenosine triphosphate (ATP) is widely considered as the energy currency in cells and plays an important role in most enzyme reactions. The concentration and consumption rate of ATP are closely related to the occurrence of many diseases, such as: hypoxia, hypoglycemia, Ischemia and Parkinson's disease etc. Traditional methods for measuring ATP include high-pressure liquid chromatography, radioisotope method, etc. Among them, high-pressure liquid chromatography is cumbersome and expensive, and the detection time needs more than 30 minutes, and the content of ATP cannot be directly measured, and the sensitivity is only at the millimolar level; radioisotope method The sensitivity is high, but the pollution is large, the protection of the sample and the waste treatment process are required, and it is ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 刘霞王振新李小梅刘殿骏
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI