Stationary liquid for preserving human saliva, preparation and application

A fixative and human body technology, applied in the field of molecular biology, can solve the problems of long storage time, inconvenient extraction, blood clot formation, etc., and achieve the effect of low cost, convenient preparation, and good preservation effect

Active Publication Date: 2012-05-09
上海泛亚基因医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The DNA usually used in PCR reactions and other molecular tests is mainly extracted from anticoagulated blood, but blood samples that have been stored for a long time will form blood clots even if sodium citrate or EDTA anticoagulants are added, which brings inconvenience to the extraction
At the same time, because blood collection can cause certain trauma to the human body and requires certain professional skills to complete, it limits the wide application of blood as a source of DNA.

Method used

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  • Stationary liquid for preserving human saliva, preparation and application
  • Stationary liquid for preserving human saliva, preparation and application
  • Stationary liquid for preserving human saliva, preparation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Saliva Genomic DNA Extraction After Fixative Fixation in Example 1

[0030] The formula of human saliva preservation fixative is: 0.25mol / L sucrose, 8mmol / L Tris-HCl (pH is 8), 3mmol / L MgCl 2 , 2.5mol / L guanidine isothiocyanate.

[0031] Preparation method of human saliva preservation fixative:

[0032] (1) First prepare 1M sucrose, 1M MgCl 2 , 6M guanidine isothiocyanate, 1M Tris-HCl (pH 8.0) and dd H 2 O, respectively by 1.1kgf / cm 2 Sterilize for 20 minutes;

[0033] (2) Taking the preparation of 100ml fixative solution as an example, mix various reagents according to the following ratio:

[0034] 1M sucrose 25ml

[0035] 1M Tris-HCl (pH 8) 0.8ml

[0036] 1M MgCl 2 0.3ml

[0037] 6M Guanidine Isothiocyanate 41.7ml

[0038] dd H 2 O 32.2ml

[0039] Total 100ml

[0040] Experimental steps:

[0041] (1) Take 500ul saliva sample (including equal volume of fixative and saliva, which has been placed at a certain temperature for a certain period of ...

Embodiment 2

[0056] Example 2 Extraction of genomic DNA from saliva samples placed at different temperatures and at different times after fixative fixation

[0057] The formula of human saliva preservation fixative is the same as embodiment 1

[0058]The preparation method of human saliva preservation fixative is the same as embodiment 1

[0059] Experimental procedure is with embodiment 1, and result is as figure 2 .

[0060] Such as figure 2 Shown: No. 1 is the fixative solution to fix human saliva, and extract genomic DNA after leaving it at room temperature (25°C) for 7 days;

[0061] No. 2 is the fixative solution to fix human saliva, and extract genomic DNA after placing it at 37°C for 7 days;

[0062] No. 3 is the fixative solution to fix human saliva, and then place it in a -20°C refrigerator for 7 days to extract genomic DNA;

[0063] No. 4 is the fixative solution to fix human saliva, and extract genomic DNA after placing it in a refrigerator at 4°C for 7 days.

[0064] T...

Embodiment 3

[0066] Example 3: Genomic DNA extraction and PCR amplification of saliva samples placed at room temperature for one month after fixative fixation

[0067] The formula of human saliva preservation fixative is the same as embodiment 1

[0068] The preparation method of the human saliva preservation fixative is the same as in Example 1.

[0069] Experimental steps:

[0070] 1. Extraction method is with embodiment 1;

[0071] 2. PCR amplification conditions:

[0072] 1) 95°C for 3 minutes

[0073] 2) 94℃30s

[0074] 3) 50℃30s

[0075] 4) 72℃30s

[0076] 5) Extend at 72°C for 5 minutes

[0077] 6) Cool at 4°C.

[0078] Step 2) to step 4) loop 35 times

[0079] Experimental results such as image 3 :

[0080] image 3 In order to use this saliva fixative to extract genomic DNA from 5 samples from different sources, there are obvious products after the above PCR amplification, which shows that the genomic DNA extracted from human saliva fixed by this fixative is suitable ...

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Abstract

The invention relates to a stationary liquid for preserving human saliva, a preparation and an application, wherein the stationary liquid for preserving the human saliva comprises the following components: sugar, Tris-HCl, MgCl2, guanidinium isothiocyanate and water. The saliva stationary liquid provided in the invention has better preservation effect; the preservation effect is obviously better than the preserving saliva with single component; and with lower cost and convenient preparation, the stationary liquid for preserving the human saliva is hopeful to become the necessary and better stationary liquid for widely using the saliva as genome DNA (Deoxyribonucleic Acid) source.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to the formula, preparation and application of a human saliva preservation fixative. Background technique [0002] In molecular biology research, DNA extraction from blood samples plays an important role in genetic engineering and molecular biology research. DNA usually used in PCR reactions and other molecular tests is mainly extracted from anticoagulated blood, but blood samples that have been stored for a long time will form blood clots even if sodium citrate or EDTA anticoagulant is added, which brings inconvenience to the extraction. At the same time, because blood collection can cause certain trauma to the human body and requires certain professional skills to complete, it limits the wide application of blood as a source of DNA. [0003] Human saliva contains exfoliated cells of the oral mucosa. Collecting saliva as a source of DNA samples has the follow...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 鲍晓妮江三多张天亮彭丽娟张健魏宁孙长胜
Owner 上海泛亚基因医学科技有限公司
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