Saliva preserving fluid and preparation method and application thereof

Abstract

The invention relates to a saliva preserving fluid and a preparation method and application thereof. The saliva preserving fluid is prepared from 1-10 mmol/L of EDTA, 1-10 mmol/L of ethylene glycol-bis(2-aminoethyl ether)tetraacetic acid, 30-70 micrograms/mL of sodium benzyl penicillinate, 30-70 micrograms/mL of streptomycin sulphate, 30-70 mmol/L of Tris-HCl, 1-25 mmol/L of 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid, 30-70 mmol/L of N-tris(hydroxymethyl)methylglycine, 5-30 micrograms/mL of protease K and 1-50 mmol/L of saccharose, and a solvent is sterile water. The saliva preservingfluid can make a DNA sample preserved in the saliva preserving fluid at the normal temperature for a long time maintain integrity through the rational combination of all the components in a specificconcentration range, and can be directly used for PCR amplification without DNA extraction; the saliva preserving fluid can be used for an NGS experiment, can adapt to complex multiple PCR experiments, and can meet the requirement long-fragment amplification.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a saliva preservation solution and a preparation method and application thereof. Background technique [0002] With the development of biotechnology, DNA samples are more and more widely used in the fields of molecular biology, medical identification, detection and treatment. The DNA usually used for PCR amplification or detection is extracted from anticoagulated blood, but blood samples are limited to a certain extent as a source of DNA, which is mainly reflected in two aspects. The extraction is inconvenient; on the other hand, collecting blood will also cause certain trauma to the human body and also rely on professional skills to complete. [0003] Compared with the traditional way of drawing blood to collect DNA, saliva DNA sample collection is a method of obtaining DNA that is harmless and painless to the human body, and is not easy to cause discomfor...

AI Technical Summary

Problems solved by technology

Saliva samples are an ideal way to obtain DNA, but inappropriate storage solutions can cause DNA degradation or contain certain components that affect downstream genetic testing

Some of the preservation solutions currently used to preserve saliva contain magnesium chloride, which can easily lead to DNA degradation. After long-term storage, it will affect the downstream long fragment amplification.

Some contain sucrose, but lack antibacterial substances. Although they can maintain cell osmotic pressure well, they are easy to cause the growth of bacteria and other microorganisms, which is not conducive to the long-term preservation of saliva.

[0005] CN105368812A discloses a saliva preservation solution and its preparation method and application. The saliva preservation solution includes the following components and contents: lithium chloride 0.1-2mol / L, Tris-HCl 1-50mmol / L, urea 0.1-2mol / L L, SDS0.1-2% (w / v), EDTA1-10mmol / L, ethanol 10-90% (v / v), the pH range of the system is between 7-10, although components such as urea and ethanol can make saliva The organic matter such as mucin and globulin in the medium is deformed, but at the same time it will also lyse the cells, which is not conducive to the integrity preservation of DNA

The saliva fixative of the present invention has a good preservation effect, and its preservation effect is obviously better than that of using a single component to preserve saliva, and the cost is low, and the preparation is convenient, but it contains protein denaturant guanidine isothiocyanate, which can inhibit nucleic acid Enzyme activity but also lyses cells at the same time, leaving DNA in a free state, which is not conducive to the integrity preservation of DNA

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