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A kind of method for measuring the content of pyrimidine dimer on DNA chain

A pyrimidine dimer, DNA chain technology, applied in the field of gene mutation detection, can solve the problems of narrow detection sample range, complex sample processing, cumbersome operation, etc., and achieve a wide sample range, good reproducibility and low detection cost Effect

Inactive Publication Date: 2011-11-30
UNIV OF SCI & TECH OF CHINA
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Problems solved by technology

[0004] The purpose of the present invention is to propose a method using the principle of immune competition to determine the content of pyrimidine dimers on the DNA chain, so as to overcome the shortcomings of the existing methods such as cumbersome operation, complicated sample processing, narrow range of detectable samples, and even environmental pollution.

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  • A kind of method for measuring the content of pyrimidine dimer on DNA chain
  • A kind of method for measuring the content of pyrimidine dimer on DNA chain

Examples

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Embodiment 1

[0012] Embodiment 1, the mensuration of pyrimidine dimer (CPD) concentration

[0013] 1. Preparation of CPD Standards

[0014] Preparation of Crude Sample of Pyrimidine Dimer

[0015] An oligonucleotide chain with only two pyrimidine bases in adjacent positions synthesized by Shanghai Shenggong Company is used. There are only two pyrimidine bases in adjacent positions in the chain. The purpose is to ensure that only A pyrimidine dimer, allowing for accurate quantification of pyrimidine dimers. The oligonucleotide chain sequence used in this example is 5'-AGA GCA GTT GACACG-3'. Place 100 μl of 150 μM oligonucleotide aqueous solution sample containing 20% ​​acetone in a glass NMR tube that cannot transmit light with a wavelength of less than 290 nm, inflate and bubble with high-purity argon gas with a purity >99.999% for 15 minutes, and seal it with a parafilm . The prepared samples were irradiated with a 300-watt high-pressure mercury lamp, the temperature of the water bath...

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Abstract

The method for measuring the content of pyrimidine dimer on the DNA chain of the present invention is characterized in that the probe DNA with a pyrimidine dimer modified at the end is fixed on the surface of the chip; the standard product of pyrimidine dimer is prepared; Mix the 80nm pyrimidine dimer standard with 0.2-5 μg / ml pyrimidine dimer antibody, and plot the response value of the surface plasmon resonance sensor to detect the mixture flowing over the chip surface against the concentration of pyrimidine dimer to obtain a standard curve; The sample is mixed with the antibody at the same concentration as the standard curve is established, and the response value of the mixture flowing through the chip surface is detected, and the concentration of pyrimidine dimer in the sample can be obtained according to the standard curve. The invention has the advantages of rapid detection, accurate quantification and good reproducibility; the chip can be reused, and the detection cost is low; the range of detectable samples is wide; Evaluate the effects of ozone layer depletion on human health.

Description

Technical field: [0001] The invention belongs to the technical field of gene mutation detection methods, in particular to a method for measuring the content of pyrimidine dimers on a DNA chain and an immunological detection method for mutations. Background technique: [0002] According to "Perfume, Fragrance and Cosmetics" (2005.6: 25-28, 32), ultraviolet radiation can cause DNA damage and form pyrimidine dimer (CPD). The formation of pyrimidine dimers can cause various diseases such as skin inflammation, skin aging, and even skin cancer. The content of pyrimidine dimer reflects the degree of DNA damage and can be used to evaluate the effectiveness of various prevention and treatment methods. Quantitative determination of pyrimidine dimer content is a necessary means to study UV damage, therefore, people have been exploring effective detection methods. [0003] Commonly used methods for measuring CPD concentration include: the radioimmunoassay (RIA) mentioned in German "Mo...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N21/55
Inventor 罗昭锋沐万孟王玉珍林琳郭晓鹏
Owner UNIV OF SCI & TECH OF CHINA
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