CN1360054A - Application of nucleic acid in antiforge purpose by gene chip technique - Google Patents
Application of nucleic acid in antiforge purpose by gene chip technique Download PDFInfo
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- CN1360054A CN1360054A CN 00127922 CN00127922A CN1360054A CN 1360054 A CN1360054 A CN 1360054A CN 00127922 CN00127922 CN 00127922 CN 00127922 A CN00127922 A CN 00127922A CN 1360054 A CN1360054 A CN 1360054A
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- nucleic acid
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Abstract
The present invention discloses the method for using nucleic acid for antiforge purpose, the process for preparing antiforge label, and the use of said method. It characterized by that the antiforge label is prepared from nucleic acid, and then checked by gene chip.
Description
The present invention is applied in the nucleic acid cryptanalysis on the concealed anti-false mark of commodity or other objects, belongs to the technical field of cryptography method.
Traditional anti-fake mark is the goods of physics or chemical property mostly, as laser hologram, optic metachromatic film, image scrambling, thermal change printing ink, magnetic stripe, software etc.Begin at present to occur the anti-fake product made of biotechnology in the world, these products mainly adopt the antigen antibody reaction principle, and are more accurate, sensitive than general anti-fake product and method.But antigen-antibody is a protein, less stable, and easy inactivation 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.
Purpose of the present invention is exactly to overcome underlinedly easily by imitated defective in false proof technical field, designs and utilizes biochip technology that nucleic acid is applied to false proof hidden mark method.
The present invention is based on the gene principle of work, proposes brand-new concealed anti-false design and technology, and its core is biomacromolecule-nucleic acid.People know that the genetic material of all life (gene) 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, and life style is varied on the formation earth.Article one, only the DNA of 1000 base pairs (1kb) promptly 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 numerous.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
5X (10
4--10
6)=10
10--10
12More than.Therefore species diversity provides inexhaustible source for anti-counterfeiting technology.Utilize the huge like this quantity of information of DNA can have the password of counting in hundreds of millions to can be used for false proof design.
The present invention is (mainly to be dna fragmentation with previously selected certain nucleic acid fragment, because of it is highly stable), matrix film has aldehyde radical on the chip because nucleotide sequence has amino side-chain, so both under proper condition (as UV-irradiation) can form with covalent linkage crosslinked, thereby nucleotide sequence is fixed on the carrier.Nucleotide sequence on the gene chip can synthetic, or directly uses the natural acid fragment, and according to design be with fluorescently-labeled oligonucleotide fragment as probe.
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.Also can the solution form add in the goods or the envelope oar is attached on the packing of goods.In order to protect on the solid phase carrier, add protective layer at the carrier surface that contains nucleic acid fragment as the nucleic acid fragment of anti-false sign.This nucleic acid concealed anti-false mark that for example available plastics film, trehalose etc. are made can be fixed on commodity or the object or on its packing material and the appurtenant.When needs are checked, anti-fake mark is taken off, throw off protective layer, dissolve attached to the nucleic acid fragment on the carrier with damping fluid.The kind of nucleic acid anti-fake mark on known detection commodity of testing staff or the object is hybridized as probe with regard to available few nucleic acid fragment at nucleic acid fragment preparation on the chip.By detecting gene chip, if can show fluorescent signal, can judge that detection commodity or object are genuine piece, otherwise puppet product or fakement for palming off.Utilize the nucleic acid password to be marked with following advantage as concealed anti-false:
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: according to the nucleic acid hybridization principle, have only when the base pair of nucleic acid on probe base sequence and the chip is complementary fully, could hybridize.Test so have only, just might detect the existence of password and the kind of password by the probe of predetermined nucleic acid encryption design.So the nucleic acid password has high specificity.
3. diversity: can be easy to design, synthetic ten million kind of nucleic acid password is as anti-fake mark, or directly uses the natural acid fragment, and is not general mutually.So the nucleic acid password can be diversified commodity and object provides unique anti-fake mark system respectively.
4. stability: the DNA password 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: the nucleic acid password 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.
Survey measuring means and be used for distinguishing that the true and false of object is the indivisible technology contents of nucleic acid cipher anti-fake method.As described above, the present invention adopts a kind of biochip technology nucleic acid to be applied to false proof method.
The invention provides accompanying drawing a: Fig. 1 and express spectrogram, strong and weak distinguishable its true and false of corresponding positions dot blot signal about pressing among two figure for nucleic acid being applied to false proof chip with biochip technology.
Embodiment: present embodiment adopts biochip technology nucleic acid to be applied to false proof method validation dna fragmentation specificity.BD-CHIP512 chip mark crossing system according to Bo Dao company carries out, and concrete steps are as follows:
1. prehybridization:
A. chip is put 2min in 95 ℃ of distilled waters baths, puts 30sec in the dehydrated alcohol after the taking-up immediately, takes out room temperature and dries;
B. get hybridizing reagent 1,2 each 6ul, put in the Eppendoff pipe of 0.5ml, put 2min in 95 ℃ of distilled waters baths, use immediately;
C. mentioned reagent is added on the chip according to posting, covered (note rifle head can not run into chip, can not leave bubble) is put in the hybridization cabin 42 ℃, 5hr.Below in the darkroom, carry out (2)-(4)
2. mark:
A. get probe 3ug at two kinds of oligonucleotide fragments of the preparation of the nucleic acid fragment on the chip,
B. see in the pipe above-mentioned two and transfer to respectively in dyestuff A and the dyestuff B pipe, add labelled reagent 1: 4ul respectively, mixing adds labelled reagent 2: 4ul, mixing behind 65 ℃ of water-bath 10min.
C. merge the liquid in above-mentioned two pipes, record mark reagent 3: 15ul adds the 250ul dehydrated alcohol, and mixing is put-20 ℃, 2hr;
D.12000rpm, 4 ℃ of centrifugal 10min precipitate standby with drying after 300ul 75% washing with alcohol.
3. hybridization
A. add hybridizing reagent 1: 6ul in sediment tube, vibration shakes up, and adds hybridizing reagent 2: 6ul, and 95 ℃ of water-bath 2min take out and place on ice rapidly behind the mixing
B. the chip through prehybridization washes to remove cover glass with distilled water, and 95 ℃ of water-bath 2min are taken out to 30sec in the dehydrated alcohol, dry;
D. probe is added to the central position in point sample district on the chip, covered, light finger are pressed and are made hybridization solution uniform distribution and slide and chip chamber not have bubble, put in the hybridization cabin, in 42 ℃ of hybridization 15hrs-29hrs.
3. develop a film
A. agent 1: the 2.5ml that will develop a film is diluted to 49ml in the beaker of 50ml, add the 1ml reagent 2 of developing a film, and beaker is put preheating in 60 ℃ of water-baths;
B. agent 3: the 2.5ml that will develop a film is diluted to 50ml in the 50ml beaker, preheating in 60 ℃ of water-baths of beaker;
C. reagent 1: the 1ml that will develop a film is diluted to 200ml in wash bottle, the chip after the hybridization washes to remove cover glass with this solution;
D. chip inserts 10min in the beaker of reagent 1,2 diluent of developing a film be preheated to 60 ℃, and moves into 10min in the beaker of reagent 3 diluents of developing a film that are preheated to 60 ℃ again;
E. take out, fully wash chip with the solution of the step 3 of developing a film, two do and are equipped with scanning
Claims (6)
1. one kind is adopted biochip technology that nucleic acid is applied to false proof method, its feature mainly is to be dissolved in thymus nucleic acid (DNA) fragment in the solution and to make it attached to being used as the concealed anti-false mark on the solid phase carrier, and utilizes biochip technology to detect the true and false.Nucleotide sequence on the gene chip can synthetic, or directly uses the natural acid fragment, is with fluorescently-labeled nucleic acid fragment as probe according to its design, and nucleic acid is made anti-fake mark.
2. the method for anti-forge cipher according to claim 1, the solid phase carrier that it is characterized in that nucleic acid fragment 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.
3. according to the method for anti-forge cipher according to claim 1 and 2, it is characterized in that directly to be fixed on any solid-state commodity, object or its packing material and the appurtenant attached to the concealed anti-false mark on the solid phase carrier.
4. the method for anti-forge cipher according to claim 1 is characterized in that the carrier surface that contains nucleic acid fragment can add protective layers such as covering plastics film or trehalose.
5. the method for anti-forge cipher according to claim 1 is characterized in that the detection method of inspection that anti-fake mark is distinguished can adopt making nucleic acid molecular hybridization.According to the nucleic acid hybridization principle, have only when the base pair of nucleic acid on probe base sequence and the chip is complementary fully, could hybridize.After both combinations, can judge whether the specificity of nucleotide sequence on the chip is correct by fluorescence on the chip.Its steps in sequence is: prehybridization, and mark, hybridization is developed a film, scanning.
6. the method for anti-forge cipher according to claim 1 or 5 is characterized in that the used probe of making nucleic acid molecular hybridization can use cy3, the cy5 dye marker.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00127922 CN1360054A (en) | 2000-12-18 | 2000-12-18 | Application of nucleic acid in antiforge purpose by gene chip technique |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00127922 CN1360054A (en) | 2000-12-18 | 2000-12-18 | Application of nucleic acid in antiforge purpose by gene chip technique |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1360054A true CN1360054A (en) | 2002-07-24 |
Family
ID=4592832
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 00127922 Pending CN1360054A (en) | 2000-12-18 | 2000-12-18 | Application of nucleic acid in antiforge purpose by gene chip technique |
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|---|---|
| CN (1) | CN1360054A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107273960A (en) * | 2017-05-25 | 2017-10-20 | 北京宝真基因科技有限公司 | Nucleotide sequence bar code substrate, antifalsification label and method for anti-counterfeit |
-
2000
- 2000-12-18 CN CN 00127922 patent/CN1360054A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107273960A (en) * | 2017-05-25 | 2017-10-20 | 北京宝真基因科技有限公司 | Nucleotide sequence bar code substrate, antifalsification label and method for anti-counterfeit |
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