Medium for adsorbing stored DNA and preparation method

A medium and mass ratio technology, applied in the field of DNA medium and preparation, can solve the problems of undescribed filter paper or fiber modification or modification

Active Publication Date: 2010-12-15
天津金歌联合生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the patent does not describe whether the filter paper or fiber has been modified or modified

Method used

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  • Medium for adsorbing stored DNA and preparation method
  • Medium for adsorbing stored DNA and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A medium for adsorbing and storing DNA is prepared by the following method:

[0029] (1) Take chitosan with a molecular weight of 56,000 and a degree of deacetylation of 85%, and prepare a 0.5% chitosan solution with a mass concentration of 1% acetic acid aqueous solution; prepare TE buffer solution (25mM Tris-HCL / 10mM EDTA, pH8.0) 500ml, preparation mass concentration is 500ml of cetyltrimethylammonium bromide aqueous solution of 5%, TE buffer solution is mixed with cetyltrimethylammonium bromide aqueous solution; A solution;

[0030] (2) Take some quantitative filter paper, soak it in the chitosan solution for 10 minutes, take it out, wash it twice with distilled water, then put it into the A solution and soak it for 10 minutes, take it out, dry it at 40-50°C, and then make a A medium that adsorbs and stores DNA.

Embodiment 2

[0032] Collection and extraction of blood DNA for STR locus typing using the medium of the present invention

[0033] Take 1 drop (about 0.2ml) of the volunteer's whole blood and drop it on a medium for absorbing and storing DNA prepared in Example 1, dry it naturally, put it in an envelope, and store it at room temperature for 7 days. mm bloody filter paper in a centrifuge tube (or in a 96-well PCR plate); add 200ul sterilized deionized water, soak for 5min, suck out the soaking solution, and repeat washing twice; dry at 100°C for 10min, wait for amplification; put in a DNA extraction tube (or wells) to construct a 10ul PCR system, and amplify for 28 cycles in the AB-9700 thermal cycler; take 1ul of the amplified product, and perform STR typing detection in the AB-3130XL genetic analyzer. The DNA typing results of 16 STR loci were obtained, and the peak heights were balanced, the map indicators were good, and there was no peak loss ( figure 1 ).

Embodiment 3

[0035] A medium for adsorbing and storing DNA is prepared by the following method:

[0036](1) be the ratio of 1: 100 by mass ratio, be that the chitosan that deacetylation degree is 85% is dissolved in the citric acid aqueous solution that mass concentration is 5% makes cationic polymer solution; By volume ratio is 1: 1 Mix the TE buffer solution with the cetyltrimethylammonium bromide aqueous solution whose mass concentration is 8% to make A solution;

[0037] (2) Immerse the wood pulp filter paper in the cationic polymer solution, soak for 6 minutes, take it out, wash it twice with distilled water, then put it into the A solution for 8 minutes, take it out, dry it, and make a kind of DNA adsorption storage medium.

[0038] The present embodiment also can select the chitosan that the deacetylation degree is 5%, the chitosan that the deacetylation degree is 50% or the chitosan that the deacetylation degree is 99% instead of the deacetylation degree is that the chitosan compo...

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Abstract

The invention discloses a medium for adsorbing stored DNA and a preparation method. The preparation method of the medium for adsorbing the stored DNA is as follows: (1) dissolving a cationic polymer in a water solution of acid the mass concentration of which is 0.1%-10% or in water to prepare a cationic polymer solution; mixing a TE buffer solution and cetyltrimethylammonium bromide (CTAB) to prepare a solution A; and (2) immersing a filter paper in the cationic polymer solution, soaking and taking out, washing by distilled water, putting into the solution A for soaking, and then taking out and drying to obtain the medium for adsorbing the stored DNA. In the invention, a protein denaturing agent is used to ensure the medium to have antibacterial activity, so that the medium can directly extract DNA from the solid medium adsorbing a biological sample or apply a PCR method for amplification, and can be used for medical diagnosis, criminal site sampling, physical evidence preservation, biological sample mailing, epidemiology detection, etc.

Description

technical field [0001] The invention belongs to the field of biological carrier production, in particular to a medium for adsorbing and storing DNA in biological samples and a preparation method. Background technique [0002] Collecting, storing, transporting and purifying nucleic acids of various biological samples at room temperature and performing PCR, SNP analysis, STR typing and restriction enzymatic hydrolysis analysis are currently used in the fields of medical diagnosis, forensic identification, and epidemiological prevention testing. Common operating techniques and methods. For the preservation and transportation of DNA in biological samples (especially blood), refrigeration is usually required, and the equipment is expensive and the operation steps are also cumbersome. What's more, since there is no effective way to prevent the DNA in the sample from being damaged, the sample cannot be stored for a long time. In addition, because some bacteria or viruses in the b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/26B01J20/32C12N15/10
Inventor 葛志强靳风民
Owner 天津金歌联合生物科技有限公司
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