Saliva preservation solution, preparation method and uses thereof

A technology for preserving fluid and saliva, applied in the field of molecular biology, can solve the problems of easy fracture, cell rupture, bacterial growth and other microorganisms, and achieve the effect of good integrity and reduced adhesion

Active Publication Date: 2016-03-02
BGI GENOMICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But, owing to containing sucrose composition, though can well keep the viscosity of this saliva preservation immobilization solution and maintain cell osmotic pressure, easily cause the growth of microorganisms such as bacteria; Magnesium chloride is as the activator of nuclease, causes the degradation of DNA easily; Guanidine thiocyanate as a denaturant can inhibit the activity of nucleases, but it is easy to cause cell rupture, making DNA exposed and easy to break

Method used

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  • Saliva preservation solution, preparation method and uses thereof
  • Saliva preservation solution, preparation method and uses thereof
  • Saliva preservation solution, preparation method and uses thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0034] The saliva preservation liquid of the present embodiment comprises following components and content: lithium chloride 0.67mol / L, Tris-HCl33mmol / L, urea 0.67mol / L, SDS0.6% (w / v), EDTA3.3mmol / L, Ethanol 30% (v / v), the system pH is 7.5.

[0035] The preparation method of the saliva preservation solution of the present embodiment is as follows:

[0036] (1) Weigh 6.036g of urea and 4.26g of LiCl, respectively, and pour them into beakers in turn (clean the beakers with pure water). Note that the weighing process should be completed in a short time as much as possible to prevent deliquescence of the samples.

[0037] (2) Measure 4.95mL Tris-HCl (1M), 9mL SDS (10%), 0.99mL LEDTA (0.5M) and 45mL ethanol respectively, pour them into a beaker in turn, stir and dissolve with a clean glass rod, and set Make up to 150mL.

[0038] (3) Filter the above preservation solution with a disposable filter membrane device, and store it in a storage bottle.

[0039] (4) Adjust the pH value ...

Embodiment 2

[0060] This embodiment studies the situation of performing PCR amplification of 10 pairs of housekeeping genes from the extracted genomic DNA, using the samples from individual 1 and individual 2 stored in saliva samples for 1 week, 2 weeks, 3 weeks, 4 weeks and After 2.5 months, the DNA extracted was used as a PCR amplification template for experiments.

[0061] The 10 pairs of housekeeping gene amplification primers used in the experiment are listed as follows:

[0062] Table 1

[0063]

[0064] The PCR reaction system is as follows:

[0065]

[0066]

[0067] The PCR reaction procedure is as follows:

[0068] 95°C for 4min; 95°C for 40s, 58°C for 30s, 72°C for 50s, 34 cycles; 72°C for 10min; 16°C∝.

[0069] figure 2 It is the agarose gel electrophoresis image obtained by PCR amplification of the saliva sample of individual 1 using the genomic DNA extracted after 2.5 months of storage in the saliva preservation solution of Example 1 as a template, and the above...

Embodiment 3

[0071] In this embodiment, low-concentration and high-concentration preservation solutions were prepared with reference to the preparation method of Example 1, and the saliva preservation solution in Example 1 was used as the medium-concentration preservation solution, and these three groups of preservation solutions were used to preserve two individuals (individual 1 and After 1 week, 2 weeks, 3 weeks, and 4 weeks, DNA was extracted from the saliva samples of individual 2), and then interrupted, random high-throughput sequencing, and comparison were performed to study the contamination of microorganisms such as exogenous bacteria.

[0072] In the present embodiment, the low-concentration storage solution includes the following components and contents: lithium chloride 0.1mol / L, Tris-HCl1mmol / L, urea 0.1mol / L, SDS0.1% (w / v), EDTA1mmol / L, Ethanol 10% (v / v), the system pH is 7.0. The high-concentration preservation solution includes the following components and contents: lithium...

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Abstract

The present invention discloses a saliva preservation solution, a preparation method and uses thereof, wherein the saliva preservation solution comprises 0.1-2 mol/L of lithium chloride, 1-50 mmol/L of Tris-HCl, 0.1-2 mol/L of urea, 0.1-2% (w/v) of SDS, 1-10 mmol/L of EDTA, and 10-90% (v/v) of ethanol, and the pH value of the system is 7-10. The saliva preservation solution of the present invention is used for preserving saliva, can maintain the integrity of DNA in cells for a long time, and can prevent bacteria and other microorganisms from breeding.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a saliva preservation solution and its preparation method and application. Background technique [0002] Nucleic acid DNA samples extracted from human tissues and body fluids are widely used in forensic identification, disease detection and treatment and other fields. At present, researchers and clinicians mainly use anticoagulant blood to extract DNA. However, agglutination occurs after anticoagulant blood is stored for a long time, which makes the extraction process inconvenient. At the same time, DNA extraction through blood collection still has the following inconveniences: (1) Professional training is required for blood collection personnel; (4) The throughput is low, and it is impossible to collect a large number of samples anytime and anywhere. [0003] In recent years, scientific research and medicine have gradually adopted saliva sampling to obtain individual...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 耿春雨李巧玲韩鸿雁叶睿蒋慧章文蔚
Owner BGI GENOMICS CO LTD
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