CN1344808A - Method of applying DNA length pleiomorphism analysis technology in nucleic acid anti-fake - Google Patents
Method of applying DNA length pleiomorphism analysis technology in nucleic acid anti-fake Download PDFInfo
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- CN1344808A CN1344808A CN 00125493 CN00125493A CN1344808A CN 1344808 A CN1344808 A CN 1344808A CN 00125493 CN00125493 CN 00125493 CN 00125493 A CN00125493 A CN 00125493A CN 1344808 A CN1344808 A CN 1344808A
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Abstract
The present invention discloses one method of applying nucleic acid to anti-fake. Nucleic acid is used to produce anti-fake mark and DNA length pleiomorphism analysis technology is used for testing truth or false. The present invention also discloses the technological process of producing anti-fake mark and the use of the process.
Description
The present invention discloses a kind of DNA of utilization length polymorphism analysis technology nucleic acid is applied to false proof method, belongs to biological field of anti-counterfeit technology.
Anti-counterfeiting technology commonly used comprises: antiforge laser holographic, magnetic anti-counterfeit, barcode are false proof, fluorescence falsification preventing, concealed anti-false, nuclear concealed anti-false, plasma concealed anti-false, UV-light is false proof, coding is false proof, temperature causes that to become false proof, computer code false proof or the like.
Traditional anti-fake mark is the goods of physics or chemical property mostly, as optic metachromatic film, image scrambling, thermal change printing ink, magnetic stripe, software etc.
Begin in the world at present with biotechnology be used in false proof on, formed various biological anti-counterfeiting technology and corresponding biological anti-fake mark.These anti-counterfeiting technologies mainly adopt the antigen antibody reaction principle, and are more accurate, sensitive than general anti-counterfeiting technology.Because antigen-antibody is protein, less stable, inactivation easily in hot environment especially, thus reduce false proof sensitivity and reliability.In addition, antigen antibody reaction is with low uncertainty, in case know wherein a kind of composition, just easily by imitated.
An object of the present invention is to overcome underlined easily, design nucleic acid DNA length polymorphism analysis technology is applied to false proof method by imitated defective in the false proof technical field of biology.
Another object of the present invention provides a kind of new anti-fake mark and detects the method for its true and false.
The invention provides a kind of new anti-counterfeiting technology and anti-fake mark, its core is a biomacromolecule---nucleic acid.The basic genetic material (gene) of all life all is a nucleic acid, and just thymus nucleic acid (DNA) or Yeast Nucleic Acid (RNA) nucleic acid are to be rearranged alternately by four kinds of bases, the difference of its permutation and combination, the diversity of life style on the formation earth.Article one, the DNA chain that contains 1000 base pairs (1kb) just has 10
603Plant permutation and combination method.Present natural biological species (animals and plants and microorganism) is more than 1,000,000 kinds, can utilize therefore that to make false proof natural biological gene innumerable.The length of each species gene level is general all greater than 10
4To 10
6Kb.If set and make anti-fake mark with the nucleic acid of 1kb length, then alternative nucleic acid password just has 10
5* (10
4--10
6)=10
10--10
12More than.Therefore comprised in the nucleic acid magnanimity information provide inexhaustible source for anti-counterfeiting technology.Utilize the huge like this quantity of information of nucleic acid keying sequence can have the password of counting in hundreds of millions to can be used for anti-fake mark.
The principle of DNA length polymorphism analysis is: adopt the different dna fragmentation of the group length thing that serves as a mark, by detecting the method that its length is identified.Though these dna fragmentation length differences but joint, two ends sequence is all identical, thereby can be according to 1 pair of public primer of joint design, with this group dna fragmentation is template, carries out pcr amplification by public primer, produces several simultaneously or tens even tens amplified fragments that vary in size.
(polymerase chain reaction is to be template with the section of DNA sequence PCR), is primer with two sections oligonucleotide, at the DNA section of specific amplification under the catalysis of archaeal dna polymerase between two sections primers in the polymerase chain reaction.
Amplified production separates through agarose gel electrophoresis, uses ethidium bromide staining then, and ultraviolet lamp is observed down and identified.
The present invention be will screening one section nucleic acid (can be DNA, or RNA), with a certain amount of (can be less to 10
-8Gram) is fixed on the solid phase carrier.Solid phase carrier can be various materials; as various natural or paper or film, porous particle or powder that regenerated fiber is made, natural or synthetic leather, plastics or various organic or inorganic polymer and resin, Paraffin Wax Semi Refined, natural or synthetic inorganic substance are as pottery, glass, crystal, metal, diatomite etc.Various printing ink and coating also can be used as the carrier of false proof nucleic acid.
In order to reach on the protection solid phase carrier as the purpose of the nucleic acid of anti-fake mark, can add protective layer at the surface of solid phase carriers that contains nucleic acid, for example available plastics film, trehaloses etc. are made protective layer.This nucleic acid anti-fake mark can be fixed on commodity or the object or on its packing material and the appurtenant.
When the needs check is told truth from falsehood; anti-fake mark of the present invention is thrown off protective layer; nucleic acid is dissolved in a certain amount of damping fluid; after adding specific restriction enzyme and reaction buffer; with this group dna fragmentation is template; by carrying out pcr amplification, produce several simultaneously or tens even tens amplified fragments that vary in size according to the public primer of 1 couple of joint design.
Amplification sheet cracked ends agarose gel electrophoresis separates, and uses ethidium bromide staining then, and ultraviolet lamp is observed down and identified.
Result and known DNA length polymorphism analysis result that nucleic acid DNA length polymorphism analysis on the anti-fake mark is gone out compare, and can judge the true and false of anti-fake mark.If the segment that both DNA length polymorphism analysis obtain, thinks then that anti-fake mark is true in size, quantitatively consistent, otherwise is false.
Utilize nucleic acid to make anti-fake mark following advantage arranged:
1. can not copying property: at first the nucleic acid password be colourless, and naked eyes are difficult to see that secondly, password is from the huge gene pool of nature, even know the password of gene but do not know to be any, the counterfeiter also can't decode and copy.And concerning the reviewer, as long as test with the analytical procedure of special use, the true and false then comes into plain view.
2. reliability: have only by the probe or the PCR primer of predetermined nucleic acid encryption design and test, just might detect the existence of password and the kind of password, so the nucleic acid password has high specificity.
3. diversity: can be easy to design ten million kind of nucleic acid password as anti-fake mark, every kind all has self corresponding probe and primer, not general mutually.So the nucleic acid password can be diversified commodity and object provides unique anti-fake mark system respectively.
4. stability: nucleic acid (particularly DNA) is stable, and particularly exsiccant DNA deposits prolongedly, is difficult for decomposing, and is suitable for the false proof of some prolonged preservation objects.
5. extensive applicability: nucleic acid only needs denier (10
-8-10
-10Gram) is present in commodity or the object and can detects, therefore be applicable to commodity and the object that kind is valuable.As scientific instrument and household electrical appliance; High-grade clothing, furniture and style, entertainment article; Senior tobacco and wine, food and nutritious prod; Rare or the important seed of agricultural and gardening; Important certificate and file; Rare cultural relics through identifying and artwork or the like, all applicable on nearly all solid-state object.
The invention provides an accompanying drawing: the information nucleic acid that Fig. 1 is obtained for DNA length polymorphism analysis technology, the segment that varies in size are the DNA of different lengths.
Just utilize DNA length polymorphism analysis technology that nucleic acid is applied to embodiment of false proof proposition below in conjunction with accompanying drawing.
Embodiment
Present embodiment is to adopt the different dna fragmentation of the group length thing that serves as a mark, by detecting the method that its length is identified.Though these dna fragmentation length differences but joint, two ends sequence is all identical, thereby can be according to 1 pair of public primer of joint design, be template with this group dna fragmentation during detection, carry out pcr amplification, produce several amplified fragments that vary in size simultaneously by public primer.(polymerase chain reaction is to be template with the section of DNA sequence PCR), is primer with two sections oligonucleotide, at the DNA section of specific amplification under the catalysis of archaeal dna polymerase between two sections primers in the polymerase chain reaction.Amplified production separates through agarose gel electrophoresis, uses ethidium bromide staining then, and ultraviolet lamp is observed down and identified that its operating process is exemplified below:
1. reacted constituent: (1) reaction buffer, (2) dna profiling, (3) special primer, (4) dNTP (comprising dATP, dCTP, dGTP and dTTP), (5) Taq enzyme.
2. reaction conditions: by 96 ℃ 30 seconds, 55 ℃ 30 seconds, 65 ℃ 3 minutes, circulate and carries out PCR for 30 times and react.
3. get 10 microlitre reaction solutions, put in 2% sepharose (containing the ethidium bromide dyestuff),, behind the electrophoresis sepharose is taken out, place and observe under 250～300nm wavelength ultraviolet lamp and photography with 100V volts DS electrophoresis.
4. identify: show the PCR product fragment of pre-sizing as shown in Figure 1, whether exist and the big or small true and false of whether correctly differentiating object with these bands.Do not show that band or stripe size, quantity is not to then being fakement.
Claims (5)
1. one kind is utilized DNA length polymorphism analysis technology that nucleic acid is applied to false proof method, it is characterized in that nucleic acid or its segment are fixed on and are used as anti-fake mark on the solid phase carrier, utilizes the DNA length polymorphism analysis technical survey true and false.
2. anti-fake mark as claimed in claim 1 is characterized in that described anti-fake mark can directly be fixed on any solid-state object.
3. anti-fake mark as claimed in claim 1 is characterized in that described anti-fake mark can add protective layers such as covering plastics film or trehalose.
4. solid phase carrier as claimed in claim 1 is characterized in that described solid phase carrier can be natural or the solid-state material of organic or inorganic such as regenerated fiber, paper, film, leather, plastics, porous particle, powder, resin, Paraffin Wax Semi Refined, pottery, glass.
5. the method for the checking security mark true and false as claimed in claim 1 is characterized in that comparing according to the DNA length polymorphism analysis result and the known nucleic acid DNA length polymorphism analysis result of nucleic acid on the anti-fake mark, can judge the true and false of anti-fake mark.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00125493 CN1344808A (en) | 2000-09-29 | 2000-09-29 | Method of applying DNA length pleiomorphism analysis technology in nucleic acid anti-fake |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00125493 CN1344808A (en) | 2000-09-29 | 2000-09-29 | Method of applying DNA length pleiomorphism analysis technology in nucleic acid anti-fake |
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| Publication Number | Publication Date |
|---|---|
| CN1344808A true CN1344808A (en) | 2002-04-17 |
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ID=4591280
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 00125493 Pending CN1344808A (en) | 2000-09-29 | 2000-09-29 | Method of applying DNA length pleiomorphism analysis technology in nucleic acid anti-fake |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102978283A (en) * | 2012-11-20 | 2013-03-20 | 天昊生物医药科技（苏州）有限公司 | Molecular internal standard quality control method and kit for biological sample nucleic acid detection |
| CN108138228A (en) * | 2015-09-29 | 2018-06-08 | 卡帕生物系统公司 | High-molecular-weight DNA sample for next generation's sequencing tracks label |
-
2000
- 2000-09-29 CN CN 00125493 patent/CN1344808A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102978283A (en) * | 2012-11-20 | 2013-03-20 | 天昊生物医药科技（苏州）有限公司 | Molecular internal standard quality control method and kit for biological sample nucleic acid detection |
| CN102978283B (en) * | 2012-11-20 | 2015-09-23 | 天昊生物医药科技（苏州）有限公司 | For Molecular internal standard quality control and the test kit thereof of biological specimen detection of nucleic acids |
| CN108138228A (en) * | 2015-09-29 | 2018-06-08 | 卡帕生物系统公司 | High-molecular-weight DNA sample for next generation's sequencing tracks label |
| CN108138228B (en) * | 2015-09-29 | 2022-05-27 | 卡帕生物系统公司 | High molecular weight DNA sample tracking tag for next generation sequencing |
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