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Composition for preserving DNA in saliva

A technology of composition and saliva, which is applied in the field of biomedicine, can solve the problems of affecting DNA stability and difficulty in fully inhibiting nuclease activity, and achieve the effects of saving processing time, facilitating nucleic acid detection and research, and reducing degradation

Inactive Publication Date: 2016-06-22
智海生物工程(北京)股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the presence of deoxyribonuclease and ribonuclease in saliva samples, which can decompose DNA or RNA, it is difficult to sufficiently inhibit the activity of nuclease at a lower concentration of surfactant, which may affect the stability of DNA

Method used

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  • Composition for preserving DNA in saliva
  • Composition for preserving DNA in saliva
  • Composition for preserving DNA in saliva

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 is used to preserve the composition of DNA in saliva

[0049] A composition for preserving DNA in saliva, its components and contents are shown in Table 1 below:

[0050] Table 1

[0051]

[0052] In the above table, % represents mass volume percentage.

[0053] The preparation method of the composition is as follows:

[0054] Weigh each component according to the ratio in Table 1, and completely dissolve disodium edetate to make a mother liquor; add other ingredients in deionized water and fully dissolve, add the above mother liquor and mix thoroughly, add Make up to final volume with water.

[0055] The composition prepared according to the above formula was fully mixed with the collected 10 saliva samples at a volume ratio of 1:1, and after being stored at room temperature for a period of time, 500 μL of each of the above mixed solutions was taken with a nucleic acid extraction or purification kit (Ningbo City Heavy Industry Co., Ltd. Ding Biotechnol...

Embodiment 2

[0060] The impact of different preservation time on the DNA in the preservation saliva of embodiment 2

[0061] The composition prepared according to the formula of Example 1 is fully mixed with the collected 2 saliva samples at a volume ratio of 1:1, respectively numbered samples 11 and 12, and these two samples are stored at room temperature for a certain period of time (respectively 0 months, 3 months, 6 months, 9 months, 12 months, 15 months), take 500 μL of each of the above mixtures and use a nucleic acid extraction or purification kit (Ningbo Zhongding Biotechnology Co., Ltd., product number: ZD-XY-Mini) to extract DNA and dissolve in 100 μL eluent respectively to obtain two corresponding samples. The DNA concentration in the sample was detected, and the results are shown in Table 3 below:

[0062] Table 3 (concentration is ng / μL)

[0063] sample

start

March

June

September

December

15th month

11

43.5

43.1

42.9

42...

Embodiment 3

[0067] In order to further characterize the influence of using the composition of the present invention to preserve saliva on DNA amplification and detection, the drug-induced deafness gene mutation detection kit was used to detect the DNA in the extracted saliva, and the DNA samples were respectively carrying the mitochondrial 12SrRNA gene 1555A>G Mutant and wild-type human genomic DNA.

[0068] The composition prepared according to the formula in Example 1 was fully mixed with the three collected saliva samples at a volume ratio of 1:1, and the samples were respectively numbered 13 (wild type), 14 (wild type), and 15 (1555A>G mutation) , stored at room temperature for one week. Take 10 μL of each of the above mixed solutions to extract DNA with the drug-induced deafness gene mutation detection kit (fluorescent PCR method) (Zhihai Bioengineering (Beijing) Co., Ltd.), and follow the sample processing of the drug-induced deafness gene mutation detection kit (fluorescence PCR me...

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Abstract

The invention relates to a composition for preserving DNA in saliva, which includes a denaturant, a buffer, a chelating agent, sodium chloride and polyethylene glycol. The composition of the present invention can effectively preserve DNA in saliva for a long time under room temperature conditions, and can effectively reduce the degradation of DNA in isolated saliva, ensure the integrity of DNA, facilitate subsequent nucleic acid detection research, and is simple to use and operate. Convenient and greatly saves the processing time of saliva samples.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to a composition for preserving DNA in saliva. Background technique [0002] At present, genetic testing has been widely used in the fields of biology, medicine, law, and medicine. Determination and location analysis, so as to obtain the genetic information of the subject. Genetic testing is of great significance for the control and prevention of genetic diseases, the formulation of individualized drug regimens, the confirmation of personal identity information, and the identification of parent-child relationships. Toxic deafness; through the detection of CYP2C19 gene polymorphisms, it is possible to predict drug administration conditions, the strength of toxic and side effects, and drug efficacy; through the detection of individual MTHFR gene C677T locus genotypes, it can prompt individuals to have stroke and other cardiovascular and cerebrovascular diseases and the risk of neonatal de...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/10
Inventor 冯东杨海艳沈东艳
Owner 智海生物工程(北京)股份有限公司
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