CN106434636A - Novel DNA steady solution and preparation method - Google Patents
Novel DNA steady solution and preparation method Download PDFInfo
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- CN106434636A CN106434636A CN201610977656.8A CN201610977656A CN106434636A CN 106434636 A CN106434636 A CN 106434636A CN 201610977656 A CN201610977656 A CN 201610977656A CN 106434636 A CN106434636 A CN 106434636A
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
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Abstract
Novel DNA steady solution is characterized by consisting of the following ingredients in composition proportion: 0.1M of potassium phosphate, 10mM of glucose and 10mM of ethylenediamine tetraacetic acid disodium salt, wherein solvent is water. The novel DNA steady solution disclosed by the invention has the advantages that blood genomes, plasmid DNA and plant genomes dissolved in the novel solution can be stored for three years at -20 DEG C or -80 DEG C and are not degraded. However, the same DNA dissolved in TE buffer solution or double-distilled water is partially degraded. According to the novel DNA steady solution disclosed by the invention, the DNA can keep steady, thereby overcoming the defect of being degraded.
Description
Technical field
The present invention relates to a kind of Novel DNA stablizing solution, belongs to biological technical field.
Background technology
DNA is that the important composition of chromosome is abundant, is Eukaryotic heredity base substance, the heredity containing species individuality
Information, can be used to build genomic library, separates required gene molecular marker related with detection etc..Due to contained by DNA
Hereditary information amount is big, and easily obtains, and with extremely stable chemical property, is that current germ plasm resource and genetic resourcess are collected
With one of the main contents for preserving.
The sequential structure of DNA molecular has three key properties：(1) Different Individual sequence is different；(2) constant throughout one's life；
(3) DNA sequence in the variant position cell of same human body is identical.Therefore DNA molecular possesses the characteristic of archives in itself, that is, have
Long-time stability, information attribute, individual specificity and can be for future reference attribute.
In fact, in the research field such as disease genomics, pharmacogenomicses, population genetics and evolution, DNA be most
Reliable evidence and the stock of research process, are widely used.While life science person is increasingly recognized that preservation DNA
For the importance of the development of systems generation from now on, various countries start to preserve DNA one after another.
DNA is to be connected the length of composition in certain sequence each other with 3 ', 5 '-phosphodiester bond by many oligodeoxynucleotide residues
Chain, the active force for forming chain is phosphodiester bond, is easily acid hydrolysis；In addition, most of DNA contain two such long-chains, two
Bar interchain is connected with hydrogen bond, hydrogen bond easy key point in sour environment, will also result in the degraded of DNA.
In order to maintain the integrity of DNA structure, it is to avoid DNA ruptures on a large scale and degrades, the optimal store method of DNA is dry
- 80 DEG C of powder or liquid nitrogen cryogenics are preserved, but will make dry powder then needs substantial amounts of DNA, difficulty of drawing materials, and are provided in actual DNA sample
Source does not have operability in collecting.And DNA is preserved with dehydrated alcohol, although subpackage can preserve in advance, but there is also and freeze repeatedly
Melt problem.And solid phase DNA Techniques of preserving is the new ideas being recently proposed, it is adaptable to persistence, and the preservation of this method
Whether substrate affects DNA subsequent experimental, preserves concrete effect, and any data of no this respect, still need time test at present.And
And for preservation during need nonexpondable sample, need to extract again DNA, complex operation, and be easily caused DNA break
Split, some test experience high to DNA prescription will be caused to carry out.
The method that existing many DNA are preserved is dissolved in TE buffer or distilled water, at -20 DEG C or -80 DEG C
Long-term preservation, As time goes on, DNA has degraded to both store methods.
In prior art, Chinese patent application CN 102559654A discloses a kind of DNA and preserves solvent, and the DNA preserves molten
The component of agent includes：Glycerol, TE buffer, 1- carboxy-N, N, N- trimethyl second lactone, wherein, the final volume of glycerol is 25%
Final concentration of 1 × TE～5 × TE, the 1- carboxy-N of～80%, TE buffer, N, N- trimethyl second lactone final concentration of 1～
6mol/L.This DNA preserves solution, and component is more, solution allocation complexity, relatively costly.
Content of the invention
It is an object of the invention to overcoming above-mentioned deficiency present in prior art, and a kind of transfection degrees of fusion height is provided,
Composition is simple, and DNA structure keeps stable, not degradable Novel DNA stablizing solution.
The present invention the adopted technical scheme that solves the above problems is：The Novel DNA stablizing solution, it is characterised in that by
Following component proportioning constitutes：
Potassium phosphate 0.1M,
Glucose 10mM,
Disodiumedetate 10mM,
Its solvent is water.
DNA storage temperature of the present invention is -20 DEG C or -80 DEG C.
The preparation method of Novel DNA stablizing solution described in one, it is characterised in that comprise the following steps：By potassium phosphate, Fructus Vitis viniferae
Sugar, disodiumedetate are proportionally added in water, and mix homogeneously.
Advantages of the present invention is：It is dissolved in poba gene group in this novel solutions, plasmid DNA and plant gene
Group, at -20 DEG C or -80 DEG C preserve 3 years non-degradable.But same DNA is dissolved in meeting Partial digestion in TE buffer, this
Invention can be very good to make DNA keep stable, solve the drawbacks of being degraded.Specifically advantage is：
1st, potassium phosphate meets the physiological range (pKa=7.2) of intracellular environment for buffer system, can keep DNA's
Stable；
2nd, glucose can increase the viscosity of solution, maintain osmotic pressure, prevent DNA from being acted on by mechanical shear stress and degrading；
3rd, disodiumedetate is called EDTA-2Na again, is a kind of good compounding ingredient in chemistry, and it has six to join
Position atom, the coordination compound of formation is called sequestration thing, its activity factor (Mg with DNA hydrolytic enzyme in body series2+) combine, so as to
Prevent the degraded of DNA molecular.
Description of the drawings
In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
Accompanying drawing to be used needed for technology description is had to be briefly described, it should be apparent that, drawings in the following description are only this
Some embodiments of invention, for those of ordinary skill in the art, without having to pay creative labor, may be used also
To obtain other accompanying drawings according to these accompanying drawings.
Fig. 1 is the agarose gel detection figure of contrast experiment 1.
Fig. 2 is the agarose gel detection figure of contrast experiment 2.
Fig. 3 is the agarose gel detection figure of contrast experiment 3.
Fig. 4 is the agarose gel detection figure of contrast experiment 4.
Specific embodiment
The present invention is described in further detail below in conjunction with the accompanying drawings and by embodiment, following examples be to this
Bright explanation and the invention is not limited in following examples.
Embodiment 1.
The Novel DNA stablizing solution of the present embodiment is made up of following component proportioning：
Potassium phosphate 0.1M,
Glucose 10mM,
Disodiumedetate 10mM,
Its solvent is water.
The 0.1M at this place refers to 1 liter of solution containing 0.1 mole of solute, and 10mM refers to 1 liter of solution containing 10 mMs of solutes.
DNA storage temperature is -20 DEG C or -80 DEG C.
Referring to Fig. 1, comparative experimentss 1：
Novel DNA stablizing solution in the present embodiment and TE solution are preserved at -20 DEG C DNA respectively right after 1 year
Than.
1% agarose gel loading 5ul such as following table
Sequence number 1-3 is that -20 DEG C of TE solution preserve DNA 1 year.
Sequence number 4-6 is that -20 DEG C of Novel DNA stablizing solutions preserve DNA 1 year.
Can be seen that by comparative experimentss 1, -20 DEG C of Novel DNA stablizing solutions preserve DNA and protect with -20 DEG C of TE solution after 1 year
After depositing DNA 1 year, the former concentration preserves DNA after 1 year, no drops substantially apparently higher than the latter, -20 DEG C of Novel DNA stablizing solutions
Solution.
Referring to Fig. 2, comparative experimentss 2：
Novel DNA stablizing solution in the present embodiment and TE solution are preserved at -20 DEG C DNA respectively right after 2 years
Than.
1% agarose gel loading 5ul such as following table
Sequence number 1-3 is that -20 DEG C of TE solution preserve DNA 2 years.
Sequence number 4-6 is that -20 DEG C of Novel DNA stablizing solutions preserve DNA 2 years.
Can be seen that by comparative experimentss 2, -20 DEG C of Novel DNA stablizing solutions preserve DNA and protect with -20 DEG C of TE solution after 2 years
After depositing DNA 2 years, the former concentration preserves DNA after 2 years, no drops substantially apparently higher than the latter, -20 DEG C of Novel DNA stablizing solutions
Solution.
Referring to Fig. 3, comparative experimentss 3：
It is right after DNA 3 years that Novel DNA stablizing solution in the present embodiment and TE solution are preserved at -80 DEG C respectively
Than.
1% agarose gel loading 5ul
Sequence number 1-3 is that -80 DEG C of TE solution preserve DNA 3 years.
Sequence number 4-6 is that -280 DEG C of Novel DNA stablizing solutions preserve DNA 3 years.
Can be seen that by comparative experimentss 3, -80 DEG C of Novel DNA stablizing solutions preserve DNA and protect with -80 DEG C of TE solution after 3 years
After depositing DNA 3 years, the former concentration preserves DNA after 3 years, no drops substantially apparently higher than the latter, -80 DEG C of Novel DNA stablizing solutions
Solution.
Referring to Fig. 4, comparative experimentss 4：
Sequence number 1-2 is that -80 DEG C of Novel DNA stablizing solutions preserve DNA 1 year.
Sequence number 3-4 is that -20 DEG C of Novel DNA stablizing solutions preserve DNA 1 year.
Sequence number 5-6 is that -20 DEG C of TE solution are preserved 1 year.
Can be seen that by comparative experimentss 4, -80 DEG C of Novel DNA stablizing solutions preserve DNA and compare -20 DEG C of Novel DNAs after 1 year
Stablizing solution preserved DNA after 1 year, and the former nucleic acid concentration is slightly higher, and at -80 DEG C, DNA is more stable, but difference is also little.
And -20 DEG C of TE solution preserves 1 year, then concentration substantially reduces, and there occurs degraded.
By above-mentioned comparative experimentss, the Novel DNA stablizing solution of the present invention, proportioning is simple, and easy to make, DNA is molten at this
Preserve in liquid so as to more stable, not degradable.
In the present embodiment in the form of comparative experimentss, OD260 refers to the absorbance of nucleic acid, for reaction solution amplifying nucleic acid
Concentration.260/280 ratio for representing the absorbance of nucleic acid and the absorbance of protein, i.e. nucleic acid purity, for estimating nucleic acid
Purity.260/230 represent the absorbance of nucleic acid with sample pure pollutant ratio, that is, desalt degree.
Embodiment 2.
The preparation method of the Novel DNA stablizing solution of the present embodiment, comprises the following steps：By potassium phosphate, glucose, second two
Amine tetraacethyl disodium is proportionally added in water, and mix homogeneously.
Above content described in this specification is only illustration made for the present invention.The affiliated technology of the present invention
The technical staff in field can make various modifications or supplement or adopt similar mode to described specific embodiment
Substitute, content without departing from description of the invention or surmount scope defined in the claims, this all should be belonged to
The protection domain of invention.
Claims (3)
1. a kind of Novel DNA stablizing solution, it is characterised in that be made up of following component proportioning：
Potassium phosphate 0.1M,
Glucose 10mM,
Disodiumedetate 10mM,
Its solvent is water.
2. Novel DNA stablizing solution according to claim 1, it is characterised in that：The DNA storage temperature is -20 DEG C or -80
℃.
3. as described in 1-2 is arbitrary Novel DNA stablizing solution preparation method, it is characterised in that comprise the following steps：By phosphoric acid
Potassium, glucose, disodiumedetate are proportionally added in water, and mix homogeneously.
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| CN201610977656.8A CN106434636A (en) | 2016-11-08 | 2016-11-08 | Novel DNA steady solution and preparation method |
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| CN201610977656.8A CN106434636A (en) | 2016-11-08 | 2016-11-08 | Novel DNA steady solution and preparation method |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101368179A (en) * | 2008-05-31 | 2009-02-18 | 安徽师范大学 | Reagent kit for extracting DNA in Chinese alligator chorion film and use method thereof |
| CN104651524A (en) * | 2015-03-13 | 2015-05-27 | 苏州新海生物科技有限公司 | Method for storing biological samples and kit |
| CN105039306A (en) * | 2015-05-29 | 2015-11-11 | 上海美吉生物医药科技有限公司 | Saliva protection agent |
-
2016
- 2016-11-08 CN CN201610977656.8A patent/CN106434636A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101368179A (en) * | 2008-05-31 | 2009-02-18 | 安徽师范大学 | Reagent kit for extracting DNA in Chinese alligator chorion film and use method thereof |
| CN104651524A (en) * | 2015-03-13 | 2015-05-27 | 苏州新海生物科技有限公司 | Method for storing biological samples and kit |
| CN105039306A (en) * | 2015-05-29 | 2015-11-11 | 上海美吉生物医药科技有限公司 | Saliva protection agent |
Non-Patent Citations (2)
| Title |
|---|
| 张惠展 等: "《DNA重组操作技术》", 31 May 2006, 上海交通大学出版社 * |
| 徐振军 等: "低温保存对绵羊精子DNA链完整性的影响", 《畜牧科学》 * |
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Application publication date: 20170222 |