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Desoxyribonucleic acid infrared spectroscopic analysis determining method

A technology of deoxyribonucleic acid and infrared spectrum analysis, applied in the field of infrared spectrum detection, can solve the problems of changing the uniformity of dry film, affecting the molecular structure of deoxyribonucleic acid, and the influence of characteristic sampling methods, etc., achieving high accuracy and high reproducibility Effect

Inactive Publication Date: 2018-08-28
CHINA AGRI UNIV
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Problems solved by technology

[0003] However, DNA is a trace substance, and its properties are easily affected by factors such as temperature, sampling method, sample volume, etc.
The repeatability, accuracy and analysis efficiency of DNA infrared spectra are different in different sampling methods; the drying temperature may affect the molecular structure of DNA, thereby affecting the repeatability and accuracy of its infrared spectra; the sample volume may change Uniformity of DNA drying film, which affects spectral repeatability and accuracy

Method used

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  • Desoxyribonucleic acid infrared spectroscopic analysis determining method
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  • Desoxyribonucleic acid infrared spectroscopic analysis determining method

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Experimental program
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Effect test

Embodiment 1-5

[0032] The infrared spectrum analysis measuring method of embodiment 1-5 calf thymus deoxyribonucleic acid

[0033] 1. The sample to be tested is calf thymus deoxyribonucleic acid sodium salt standard (Sigma, USA). Accurately weigh X μg of deoxyribonucleic acid and dissolve it in 150 μL of nuclease-free water, mix well, and prepare Y μg / μL of deoxyribonucleic acid solution.

[0034] 2. In order to ensure the reliability of the sample, the purity of the calf thymus deoxyribonucleic acid solution was identified by an ultra-trace ultraviolet spectrophotometer (Midrop, China), and A260 / A280 was 1.8, indicating that the purity of the calf thymus deoxyribonucleic acid solution was high.

[0035] 3. Take 15 μL calf thymus deoxyribonucleic acid assay solution and drop it on a 96-well silicon plate, and dry it at Z°C for 0.5 h to form a smooth film.

[0036] 4. Scan with a Fourier transform infrared spectrometer with a high-throughput screening attachment (HTS-XT), where the scanning ...

Embodiment 6

[0043] Embodiment 6 deoxyribonucleic acid infrared spectrometry key factor analysis

[0044] 1. Infrared spectroscopy measurement and analysis of different sampling methods

[0045] Infrared spectrum sampling methods were potassium bromide tablet method, micro-infrared point mode, micro-infrared imaging mode (Spotlight 400, PerkinElmer Inc, USA) and HTS-XT method. All acquisitions were in transmission mode.

[0046] Potassium bromide compression method: Calf thymus deoxyribonucleic acid fibrous sample and potassium bromide in a ratio of 1:100, take 0.5 mg calf thymus deoxyribonucleic acid sample and 50 mg KBr sample, mix and grind, compress into tablets, and scan by infrared spectroscopy. Spectral scanning range 4000cm -1 -400cm -1 , resolution 4cm -1 , the cumulative number of times is 32. Three parallel runs were performed and the average spectrum was taken.

[0047] Take 15 μL of 0.5 mg calf thymus deoxyribonucleic acid solution, form a dry film in an oven at 30°C, and ...

Embodiment 7

[0065] Embodiment 7 Salmon deoxyribonucleic acid infrared spectroscopic analysis and determination

[0066] 1. The sample to be tested is salmon thymus deoxyribonucleic acid sodium salt standard (Sigma, USA). Accurately weigh 1000 μg of deoxyribonucleotide and dissolve it in 150 μL of nuclease-free water, mix well, and prepare 6.667 μg / μL of deoxyribonucleic acid solution.

[0067] 2. In order to ensure the reliability of the sample, the purity of the salmon deoxyribonucleic acid solution was identified by an ultra-micro ultraviolet spectrophotometer (Midrop, China), and the A260 / A280 was 1.8, which indicated that the salmon deoxyribonucleic acid solution had a high purity.

[0068] 3. Take 15 μL salmon DNA assay solution and drop it on a 96-well silicon plate, and dry it at 30°C for 0.5h to form a smooth film.

[0069] 4. Scan with a Fourier transform infrared spectrometer with a high-throughput screening attachment (HTS-XT), where the scanning range is 4000cm -1 -400cm -1...

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Abstract

The invention provides a desoxyribonucleic acid infrared spectroscopic analysis determining method, which comprises the following steps that 1, nuclease-free water is used for preparing a desoxyribonucleic acid solution; an ultraviolet spectrophotometer is used for determining that the A260 / A280 of the desoxyribonucleic acid solution is 1.8 to 2.0; the desoxyribonucleic acid solution is used as asample to be tested; 2, a solution of the sample to be tested is dropped on a micropore plate; drying is performed until a thin film with the flat surface is formed in a micropore; 3, a fourier infrared spectrometer with a high-flux screening accessory HTS-XT is used for scanning; the average spectrum is solved; 4, the spectrum of the sample to be tested and the existing desoxyribonucleic acid standard spectrum diagram are compared; the sample to be tested is identified according to the similarity of the characteristic peak. The desoxyribonucleic acid infrared spectroscopic analysis determining method combining with analysis efficiency and analysis accuracy is provided; the spectrum collection of 120 samples can be completed within 2h; the method is a desoxyribonucleic acid detecting method with high efficiency, high repeatability and high accuracy.

Description

technical field [0001] The invention relates to the technical field of infrared spectrum detection, in particular to a deoxyribonucleic acid infrared spectrum analysis and determination method. Background technique [0002] Deoxyribonucleic acid carries genetic information, plays an important role in the growth, development and reproduction of organisms, and can be used in many fields, such as cancer diagnosis, drug treatment and species identification. Generally speaking, traditional DNA research methods are based on DNA sequences, which are time-consuming and rely on experienced operators and known specific sequence information. Therefore, it is necessary to develop a simple, rapid and reliable method for analyzing DNA. The base sequence of deoxyribonucleic acid determines the specificity of organisms, and its diversity also constitutes the diversity of composition and structure of deoxyribonucleic acid. Therefore, differences in the composition and structure of deoxyrib...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/3563G01N1/40
CPCG01N1/4022G01N21/35G01N2001/4027G01N2021/3595
Inventor 刘贤韩娅红高冰韩鲁佳姚玉梅
Owner CHINA AGRI UNIV
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