Free DNA preservation reagent and preparation method thereof

A technology for storing reagents and deionized water, which is applied in the fields of life science and biology, can solve the problems of increased transportation costs, optimal within an hour, degraded within 6 hours, increased content after 24 hours, and detection failure, etc., to achieve amplification best efficiency

Pending Publication Date: 2021-04-16
南昌艾迪康医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The size of these free DNA fragments is concentrated in 100-300bp. According to many literature reports, generally EDTA anticoagulant tubes need to separate plasma as soon as possible after blood collection. It is best within 2 hours, degraded after 6 hours, and the content increases after 24 hours. It is because the cell-free DNA released after apoptosis in the blood collection tube increases, which will dilute the original cell-free DNA, especially for non-invasive production screening, the cell-free

Method used

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  • Free DNA preservation reagent and preparation method thereof
  • Free DNA preservation reagent and preparation method thereof
  • Free DNA preservation reagent and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Prepare free DNA preservation reagents according to the following mass percentages:

[0021]

[0022] Preparation method: put disodium edetate, Tris, sodium chloride, and glycine in a beaker, add 80% deionized water to dissolve, then add formaldehyde, adjust the pH to 5.8-6.8 with 1% hydrochloric acid, and finally add the remaining Dilute to 100mL with deionized water, mix well; filter and sterilize with 0.25μm filter membrane to obtain free DNA preservation solution.

Embodiment 2

[0024] Using the cell-free DNA preservation solution prepared in Example 1, the peripheral blood of pregnant volunteers was collected, and the whole blood cell-free DNA was stored at room temperature for 5 days. Using the above-mentioned whole blood, use Annoroad nucleic acid extraction reagent and fetal chromosome aneuploidy (T21, T18, T13) detection kit to extract cfDNA and build a library. The results and library fragment analysis are as follows:

[0025]

[0026]

[0027] figure 1 It is the peripheral blood of a pregnant woman preserved with the preservation solution of the present invention for 5 days. The fragment analysis chart of the library shows that the fragment size conforms to the ratio of 200bp-300bp fragment / 200bp-400bp fragment>70%, and the position of the main peak is <280bp.

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Abstract

The invention discloses a free DNA preservation reagent which comprises ethylenediamine tetraacetic acid potassium salt (EDTA (ethylene diamine tetraacetic acid) salt), formaldehyde, glycine, Tris- HCL, sodium chloride and deionized water. The cleaning agent comprises the following components in percentage by mass: 1.5-5% of the ethylenediamine tetraacetic acid potassium salt, 0.1-1% of the formaldehyde, 1-3% of the glycine, 5-10% of the Tris-HCL, 1-20% of the sodium chloride and the balance of deionized water. In addition, the invention also discloses a preparation method of the free DNA preservation reagent. The free DNA preservation reagent disclosed by the invention can be used for preserving complete DNA fragments and can be preserved for at least 3-5 days at room temperature, so that the preservation and transportation costs are greatly reduced.

Description

technical field [0001] The invention belongs to the field of life science and biotechnology, in particular to a free DNA preservation reagent and a preparation method. Background technique [0002] Circulating free DNA (cfDNA) is extracellular DNA fragments discovered in 1948, mainly derived from cell necrosis and apoptosis. The cfDNA released by tumor cells is usually called circulating tumor DNA (ctDNA), some of which can reflect tumor-specific characteristics, such as somatic mutations, etc. By detecting ctDNA, it can well represent the characteristics of the patient's tumor, and dynamically monitor the changes of ctDNA. The patient's disease recurrence and curative effect were evaluated. With the discovery of cell-free fetal DNA (cffDNA) in maternal blood, non-invasive prenatal screening (non-invasive prenatal) has been developed. The size of these free DNA fragments is concentrated in 100-300bp. According to many literature reports, generally EDTA anticoagulant tubes ...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 孙翠莲
Owner 南昌艾迪康医学检验实验室有限公司
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