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Salivary DNA preservation liquid and preparation method and use thereof

A technology for preserving liquid and saliva, applied in the field of molecular biology, can solve problems such as corruption, complex composition, bacterial growth, etc., and achieve the effects of ensuring stability, enhancing solubility, and simple formula

Pending Publication Date: 2019-04-09
北京中科卓明生物医学研究所有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the components of saliva mainly include water, oral mucosa cells, bacteria, salivary amylase, viscous sugar and other substances. The components are complex and difficult to store for a long time. Therefore, it is very important to avoid saliva corruption and maintain the integrity of genomic DNA in exfoliated cells.
[0004] The protective agent composition in the existing patent is complex, and some of them are not conducive to the preservation of DNA integrity, such as patent publication numbers CN102440234B and CN105368812A composition is very complex; CN102919218B contains magnesium chloride, magnesium ion is a nuclease activator, which is not conducive to the stability of DNA Preservation of property and integrity; CN102919218B and CN105039306A respectively contain sucrose and glucose components, although the viscosity of the saliva preservation fixative can be well maintained and the cell osmotic pressure is maintained, but it is easy to cause the growth of microorganisms such as bacteria; magnesium chloride is used as a nuclease activator, which can easily cause DNA degradation

Method used

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  • Salivary DNA preservation liquid and preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Preparation of saliva DNA preservation solution

[0029] The saliva DNA preservation solution of this embodiment includes the following components and contents: SDS 1.5% (w / v).

[0030] The preparation method of the saliva DNA preservation solution of the present embodiment is as follows:

[0031] a. Weigh 0.75g of SDS into a 50ml beaker, add 40ml of ultrapure water, and mix evenly with a magnetic stirrer;

[0032] b. Transfer the SDS solution in the beaker to a 50ml volumetric flask, and dilute to the mark with ultrapure water;

[0033] d. Filter the above preservation solution with a disposable filter membrane device and store it in a storage bottle.

[0034] Or optionally, wherein the concentration of the SDS storage solution can be prepared as 10% (w / v) for later use; when preparing, accurately measure the SDS storage solution, and then use sterile water to make it volume; the resulting solution is operated according to step (d) to remove Bacteria; step...

Embodiment 2

[0035] Example 2 Collection and preservation of saliva samples

[0036] In this embodiment, the saliva sample collection and storage methods are as follows:

[0037] (1) Rinse your mouth 30 minutes before sampling to remove food residues and residual microorganisms, and do not eat, drink, brush your teeth, smoke or chew gum within 30 minutes after rinsing your mouth;

[0038] (2) Before spitting, relax the cheeks and rub gently for 15-30 seconds to produce saliva, and collect the saliva into a sterile 2mL EP tube;

[0039] (3) Collect 2 mL of a person's saliva according to the above method;

[0040] (4) Get the saliva DNA preservation solution prepared in Example 1 equal to the volume of saliva and add it to the saliva sample, and mix it upside down for 10-20 times;

[0041] (5) After mixing, the sample can be stored at room temperature.

Embodiment 3

[0042] Example 3 DNA extraction from saliva samples

[0043] The extraction method of saliva sample DNA in the present embodiment is as follows:

[0044] (1) Take 500ul of saliva during storage, add 1ml buffer (100mmol / L Tri-HCl, pH8.0, 0.5% SDS, 10mM EDTA) and 6ul 20mg / ul proteinase K, shake and mix on a vortexer;

[0045] (2) The saliva treated in the above step (1) in this embodiment was treated in a water bath at 55°C for 20 minutes;

[0046] (3) Add 600ul phenol: chloroform: isoamyl alcohol (25:24:1), mix by inverting, centrifuge at 10000rpm for 5min, and take the supernatant;

[0047] (4) Add 600 ul of chloroform:isoamyl alcohol (24:1) to the supernatant of the above step (3) in this embodiment, centrifuge at 10,000 rpm for 5 min after inverting and mixing, and take the supernatant;

[0048] (5) Add an equal volume of isopropanol pre-cooled at -20°C to the supernatant of the above step (4) in this example, mix well and precipitate for 1 hour;

[0049] (6) Centrifuge t...

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Abstract

The invention discloses salivary DNA preservation liquid, which is prepared from the following component in the following content: 0.1 to 8 percent (w / v) of SDS. The salivary DNA preservation liquid disclosed by the invention is simple in formula, and is used for preserving salivary DNA samples of a human body and can preserve the salivary DNA for a long time at room temperature, and the DNA in the preserved saliva is high in integrity; the salivary DNA preservation liquid can ensure the stability of transportation of a saliva sample between various collection sites and is applicable to gene detection and relevant scientific researches.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a saliva DNA preservation solution and its preparation method and application. Background technique [0002] DNA samples are widely used in the fields of molecular biology, medical identification, detection and treatment, and occupy an important position. 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. On the one hand, blood is easy to form agglutination after long-term storage. 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] Saliva is a body fluid that is easily obtained by the human body. It is more convenient to collect and non-invasive. It is an ideal body fluid sample and suitable ...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/10
Inventor 郝淑静
Owner 北京中科卓明生物医学研究所有限公司
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