CN102643912B - Amplification primer for detecting mink derived ingredients - Google Patents
Amplification primer for detecting mink derived ingredients Download PDFInfo
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- CN102643912B CN102643912B CN201210105454.6A CN201210105454A CN102643912B CN 102643912 B CN102643912 B CN 102643912B CN 201210105454 A CN201210105454 A CN 201210105454A CN 102643912 B CN102643912 B CN 102643912B
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Abstract
The invention relates to an amplification primer for detecting mink derived ingredients and belongs to the detection of special economic animal mink derived ingredients. The amplification primer has nucleotide sequences that forward primers are 5'-CTTCAACCTCAACATCATCACC-3', and reverse primers are 5'-GACATACATTGTATTCATTCTAAGCG-3'. The simplicity, the species specificity and the high-degree sensitivity of a species specificity polymerase chain reaction (PCR) method are realized, and when the method is used, the detection of animal derived ingredients in food, feed and Chinese medical herbs is greatly improved and is very convenient. Compared with other molecular biology techniques, the amplification primer has the advantages that the price is lower, the speed is higher, and the amplification primer is more suitable for the ordinary detection of most samples.
Description
Technical field
The present invention relates generally to the detection of special economic animal mink derived component, relates in particular to Auele Specific Primer and the test kit of mink gene tester.
Background technology
Mink belongs to Mammalia, Carnivora, Mustelidae, weasel genus (Mustela) in zoological taxonomy, 2 kinds of american mink (Mestela vision) and European minks (Mustela lutreola) are arranged under wild state, countries in the world artificial breeding at present is the descendant (Wang Jiyuan, 1990) of american mink for the mink that obtains fur.China carries out the relevant breeding technology research of furbearer and starts from the fifties in last century, the special economic animal resource characteristics unique with it, important economic worth and the growing market requirement and the emerging specialty industries of China's agricultural of becoming.China is that special economic animal is raised big country, and only the furbearer livestock on hand just reaches 3.28 hundred million.Mink skin, fox skin and Persian lamb skin become three large pillar products of international fur coat trade already.
At present, food-safety problem has caused showing great attention to of national governments and human consumer, and esp meat food, animal-feed security etc. concerns that the problem of people ' s health, common people's vital interests has obtained extensive attention.The thing that spreads and be transmitted to the mankind all over the world such as mad cow disease, bird flu, rabies, canine distemper also has generation, and the discriminating of the animal derived materials of livestock and poultry meat product at present detects the fields such as industry, foreign trade, market and catering trade that related to meat product.Inspection and quarantine department is day by day strong to the demand that detects the new and high technology progress, particularly outstanding in aspects performances such as feed trade, import and export epidemic prevention inspection, the supervision of meat product consumption market.Ermine meat, as the fittings of aquaculture, often is used as feed and meat product by illegal retailer, does not even pass through the inspection and quarantine program, once carry virulence factor, has greatly endangered human consumer's health, has upset livestock product and meat product market order.Simultaneously, the fur products consumption market belongs to high-end article of consumption, in interests, drives, and the phenomenon of adulterating, mix the spurious with the genuine in fur products market happens occasionally, the imitated fur products had, with the naked eye or usual way be difficult to identify the true and false, reap staggering profits thus.
Mink is a kind of as special economic animal, fur is the starting material of producing all kinds of high-grade fur productses, the ermine heart can be used as medicine, for the treatment rheumatic heart disease, certain effect is arranged, product has been arranged at present as market sales such as sharp heart balls, ermine meat, generally as animal-feed, also is processed in recent years food and uses in addition.For there being following several phenomenons on market, be necessary to develop a kind of quick, effective ways of mink source Components identification.Along with the progress of fur coat processing means and fur nitre dyeing technique, be difficult to distinguish the fur true and false according to the sense organ range estimation, interests pretend to be the phenomenon of mink fur to happen occasionally with artificial fur, other cheap animal skins under driving; In addition, when the ermine heart is used as medicine, also there is illegal businessman to pretend to be the phenomenon of the ermine heart with heart, the rabbit heart etc.; In recent years, mutton roll market is more chaotic, ermine meat, fox meat, recoon dog meat and other meat are adulterating in the mutton roll of a lot of compactings, from sense organ, be difficult to differentiate, therefore, this research emphatically, from identifying the angle of mink derived component, develops the detection kit on a kind of molecular level, have highly sensitive, specificity good, the advantage such as easy to operate.Work out a kind of fowl poultry kind animal derived materials of differentiating accurately, fast, reliably, comprise the method for identifying special economic animal mink derived component, just be necessary very much.
At present, in order to determine the true composition of food and feed, developed the method for much animal derived materials being differentiated, the methods such as physics, chemistry, immunology and molecular biology have been arranged.In molecular biology method, molecular marking technique demonstrates huge potentiality to be exploited and wide application prospect with its advantage such as quick, accurate, stable, efficient.Differentiate that at the animals and plants derived component molecule marker of application in detection mainly comprises core DNA, RNA, Mitochondrial DNA (mtDNA) and protein molecular marker etc. a few days ago.The PCR molecule marker is differentiated detection technique, there is specificity high, with strong points, highly sensitive, easy fast, lower to detecting sample requirement, such as being no matter through way far away transportation or cryopreservation outmoded sample for many years, perhaps after certain pressure, temperature processing, as long as can extract genomic nucleic acids, just may be used to the PCR reaction.
Mammals and bird mtDNA are relatively independent in heredity, be double-stranded superhelix ring molecule, there is genome little (about 16kb), there is no tumor-necrosis factor glycoproteins, contain in inorganization specificity, each cell in biont the characteristics such as a large amount of Mitochondrial Genome Overview (the about 1000-2300 of Mammals copy).Therefore, differentiate that with the mtDNA molecule marker livestock and poultry meat product and feed animal derived materials compare with the core DNA molecular marker, have highly sensitive, tolerance range good, quick, degrade little (in the course of processing, mtDNA keeps more complete), stablize the advantages such as easy to operate.The These characteristics of the PCR molecular marking technique based on animal mtDNA, therefore differentiate that at animal derived materials the application in detecting has broad prospects.
External the derived component of ox, sheep, pig, chicken is studied to report, domestic the derived components such as ox, sheep, pig, chicken, donkey, horse, deer genus, lion, tiger, leopard are identified and are studied report.In the research with molecular biology method identification and detection animal derived materials, the research that discriminating detects to the mink derived component seldom, the people such as Zhang Lihua utilize the method that enzyme is cut to study the analysis of ermine heart plastosome mtDNA identification and characterization, adopt DNA restriction fragment length polymorphism (mtDNA-RFLP) analytical technology and polymerase chain reaction (PCR) amplifying cells pigment b Gene Partial sequence fragment.Result shows that fresh ermine heart tissue can extract complete mtDNA, and can amplify the Cytb gene of 310bp size, variant with the dirty mtDNA restriction endonuclease map of heart.But the RFLP method has its advantage will choose cheap effectively applicable enzyme.The people such as Tu Jianfeng have studied from the angle when animal species classification the Phylogenetic that 12 S rRNA Gene of Jinzhou-Mink sequencing and weasel belong to animal.The people such as Zhang Honghai, to the structural analysis of mustelid mitochondrion DNA control area, further do not do the detection of derived component.Due to simplicity, species specificity and the high sensitivity of species specificity PCR method, the use of the method greatly improves and has facilitated the detection of animal derived materials in food, feed and Chinese medicinal materials.With other Protocols in Molecular Biologies, as sequential analysis, PCR-RFLP or RAPD (RandomAmplified Polymorphic DNA), compare, species specificity PCR more cheaply, more fast, be more suitable for the conventional sense of most of samples.Based on above-mentioned advantage, the researchist more and more tended to use this technology in recent years.
Summary of the invention:
The invention provides a kind of amplimer that detects the mink derived component.
The scheme that the present invention takes is:
Detecting the amplimer nucleotides sequence of mink derived component classifies as:
Forward primer: 5 '-CTTCAACCTCAACATCATCACC-3 ';
Reverse primer: 5 '-GACATACATTGTATTCATTCTAAGCG-3 '.
When amplimer of the present invention and mink genome carry out pcr amplification reaction, the nucleotide sequence of its amplified production is as described in SEQ ID No.1.
The amplimer of detection mink derived component of the present invention is for the preparation of test kit.
The present invention adopts amplimer to carry out pcr amplification to mink DNA sample, and detects amplified production.
The present invention detects the method for mink derived component, and the nucleotide sequence with amplified production is the mink sample as the described sample of SEQ ID No.1.
The present invention detects the method for mink derived component, and its PCR response procedures is: denaturation: 94 ℃, and 3min; Enter circulation: 94 ℃ of sex change 60s, 66 ℃ of annealing 60s, 72 ℃ are extended 60s, 35 circulations; Stop extending: 72 ℃, 7min; Preserve reaction product for 4 ℃.
Advantage of the present invention is: have simplicity, species specificity and the high sensitivity of species specificity PCR method, the use of the method greatly improves and has facilitated the detection of animal derived materials in food, feed and Chinese medicinal materials.With other Protocols in Molecular Biologies, as sequential analysis, PCR-RFLP or RAPD (RandomAmplified Polymorphic DNA), compare, the present invention more cheaply, more fast, be more suitable for the conventional sense of most of samples.
The accompanying drawing explanation
Fig. 1 is the electrophoresis detection figure as a result of genomic dna;
M:marker in figure, 1: mink 2: blue fox 3: silver fox 4: recoon dog 5: pig 6: chicken 7: dog 8: sheep;
Fig. 2 be 6 pairs of primers respectively with mink Genomic PCR amplification figure;
M:marker in figure, 1-6:marker1-marker6 carries out PCR with the mink genome respectively and reacts, the electrophoresis detection result;
Fig. 3 is that Auele Specific Primer is for Genomic PCR amplified production electrophorograms such as minks;
M:marker in figure, 1: mink 2: blue fox 3: silver fox 4: recoon dog 5: pig 6: chicken 7: dog 8: sheep
Fig. 4 is sensitivity test test electrophorogram;
M:marker in figure, 1: mink genome original content; 2-6: concentration dilutes 50%, 10%, 1%, 0.5%, 0.1% successively.
Embodiment
The PCR detection kit of embodiment 1 mink source components D NA and the foundation of detection method
One, the PCR of mink source DNA detects the design of primer
Retrieve 39 of Carnivora mustelid mtDNA sequence relevant informations from GenBank, comprising weasel belongs to, ermine belongs to, Lutra etc., this research is with reference to the sable of announcing in GenBank, wolverine, the Japan ermine, blue fox, silver fox, the animal mitochondria genome sequences such as racoon dog, sequence is carried out to the blast sequence alignment, species conserved sequence according to sable, analyze the specificity site, utilize primer-design software primer5.0 to design and differentiate that the PCR Auele Specific Primer that detects the mink derived component can only amplify mink DNA specific fragment, other animal DNA restriction fragment can not increase.
1, experiment in earlier stage, based on mentioned above principle, six pairs of primers have been designed, name respectively mink01 to mink06, and will design sheep in primer and GenBank, pig, chicken mtDNA sequence compares, the designed primer of preliminary judgement is only for ermine, to have specificity, and primer entrusts the handsome corporate agent in Shanghai synthetic.F (Forward): represent forward primer, R (Reverse): represent reverse primer.
2, primer has carried out a series of primer specificity checking amplification tests after synthesizing, and gropes suitable amplification temperature and reaction system condition, finally determines a best primer of specificity, as the primer of test water ermine source property check.Mink blood sample genome extracts, and adopts blood Genome DNA Pxtraction Kit test kit to carry out the extraction of genomic dna, purchased from precious biotechnology (Dalian) company limited, article No. D9081.6 pairs of primers are carried out to pcr amplification reaction with the mink genome respectively, with agarose gel electrophoresis, detect amplification, see Fig. 2.Experimental result shows, the 5th pair of primer has stronger specificity, can be used as the Auele Specific Primer that the mink source sexual element detects.Primer sequence is as follows: forward primer MinkF (22bp): 5 '-CTTCAACCTCAACATCATCACC-3 '; Reverse primer MinkR (25bp): 5 '-GACATACATTGTATTCAT
TCTAAGCG-3 '; Pcr amplification product is delivered to Shanghai and give birth to work biotechnology company limited and carry out sequencing, extension increasing sequence length is 343bp, and sequence results is as described in SEQ ID No.1.
This sequence and Genbank download sequence are carried out to blast and compare, corresponding sequence similarity degree reaches 99%, and further illustrating this amplified fragments is the mink source specific sequence.
Two, the structure of the PCR detection kit of mink source DNA
At the above-mentioned Auele Specific Primer obtained on right basis, be designed for the test kit that the PCR of mink DNA detects, test kit comprises detection solution, this detects in solution containing 10 * PCR damping fluid, 10u mol/L forward primer: 10umol/L
Reverse primer: 10u mol/L dNTP:5U/uLTaqDNA polysaccharase; Wherein, described forward primer sequence is MinkF (22bp): 5 '-CTTCAACCTCAACATCATCACC-3 '; Reverse primer MinkR (25bp): 5 '-GACATACATTGTATTCATTCTAAGCG-3 ';
Three, the foundation of the PCR detection method of mink source DNA
1, the preparation of DNA sample
The DNA sample preparation methods of the present embodiment, with reference to " DNA and RNA basic experiment technology " (Sheng little Yu etc. translate for Science Press, A.J. Kazakhstan Wood chief editor), is specially:
(1) sample preparation:
Muscle samples: take the muscle tissue chopping of about 100g mink, spend the night in 120 ℃ of bakings, be ground into powder with tissue grinder.
Blood sample: blood sample adds 2% disodium ethylene diamine tetraacetate (EDTA) anti-freezing, freezing preservation.
Feed powdered sample: can be directly used in the extraction of DNA.
(2) DNA extraction
Blood class test sample, adopt blood Genome DNA Pxtraction Kit test kit to carry out the extraction of genomic dna, purchased from precious biological above journey (Dalian) company limited, article No. D9081.Other organizes class and bone meal class sample to adopt animal derived plant feed genome DNA extracting reagent kit to be extracted, purchased from TIANGEN Biotech (Beijing) Co., Ltd., and article No. DP323-03.Fig. 1 is shown in the genome dna electrophoresis check: 1: mink 2: blue fox 3: silver fox 4: recoon dog 5: pig 6: chicken 7: dog 8: sheep
2, PCR reaction
(1) reaction system:
Get 2ul testing sample DNA solution, add the solution 6.2ul of the test kit detected for the regular-PCR of mink DNA, then add the sterilizing ultrapure water to cumulative volume 25uL; Above each component is added in 0.2mL PCR reaction tubes to 5000r/min, centrifugal 10s; Then carry out pcr amplification by following conditional parameter.
(2) reaction conditions:
Denaturation: 94 ℃, 3min;
Enter circulation: 94 ℃ of sex change 60s, 66 ℃ of annealing 60s, 72 ℃ are extended 60s, 35 circulations;
Stop extending: 72 ℃, 7min;
Preserve reaction product for 4 ℃.
3, result judgement
The amplified production electrophoresis result is shown in (Fig. 2), and with D2000bp marker contrast, in the position of bp more than 400, the size place occurs that the specific amplification band is positive, does not have the specific amplification band negative.
Principle:
(1) sex change of template DNA: template DNA, after being heated to 94 ℃ of 5min, dissociates the double-stranded or double-stranded DNA that form through pcr amplification of template DNA, makes it to become strand, so that it is combined with primer, for next round, reacts and prepares;
(2) renaturation of template DNA and primer: template DNA is after heat denatured becomes strand, and temperature is down to 57 ℃, the complementary sequence pairing combination of primer template DNA strand;
(3) extension of primer: DNA profiling one primer binding substances is under the effect of Taq archaeal dna polymerase, take dNTP as reaction raw materials, the target sequence DNA sequence dna is template, by base pairing and semiconservative replication principle, synthetic new and the semiconservative replication chain complementation of template DNA chain, recirculation sex change one renaturation one is extended three processes, just can obtain more " semiconservative replication chain ", and this new chain can become again the template of circulation next time.
4, amplification mink DNA product sequencing sequence:
The mink pcr amplification product is carried out after purifying serving sea gives birth to work biotechnology company limited and is checked order.The sequencing result of PCR specific amplification products is as follows, and sequencing result is through showing with the GenBank compare of analysis, and this sequence fragment reaches 99% with the D-loop ring section homology of the sable mt-DNA delivered.
Mink PCR product sequencing sequence result is as described in SEQ ID No.1.
The specific detection experiment of the PCR detection method of embodiment 2 mink DNA to ermine derived component DNA in meat and blood sample
For the phenomenon of with mink meat, pretending to be mutton roll there will be in market, this test detects the mink derived component from the angle of primer amplified fragment, to providing a kind of detection means for detecting in mutton roll the mink meat that whether adulterated, for quality testing department provides a kind of Molecular Detection means.Bought mutton, beef, chicken in market and extracted genome standby.Muscle tissue is extracted DNA and is adopted above-mentioned test kit to complete.Adopt the detection kit that contains Auele Specific Primer and the detection method of embodiment 1 to detect mink DNA sample.Detect other furbearer DNA samples such as blue fox, racoon dog simultaneously, assessed with the specificity of this method.
Test-results shows: the designed primer of a present invention specificity amplification primer to mink DNA, there is no the specific amplification band for derived component genomic dnas such as the sheep related in test, ox, blue fox, recoon dog, chickens, can reach the effect that detects the mink derived component.
The specificity experiment of the PCR detection method of embodiment 3 mink derived components to DNA in fur
Fur products is luxury goods, under interests are ordered about, being no lack of someone adulterates even and to use the personation fur imitation to pretend to be mink fur, perhaps with rabbit, dogskin etc., serve as mink skin, be difficult to identification through the common naked eyes of a series of processing clear, have only by further laboratory facilities and can effectively differentiate differentiation.This experimental design is as follows, choose several animal furs, adopt the method for extracting genomic dna in fur to extract DNA, fur, raw fox skin, recoon dog skin, dogskin and rabbit genomic dna have been obtained respectively, adopt test kit and method in embodiment 1 to carry out the specific band check, experimental result shows that the provable primer of the above results is only special to mink.
The PCR detection method sensitivity experiment of embodiment 4 mink DNA
The mink genomic dna of preparation is carried out to gradient dilution, is diluted to respectively 100%, 50%, 10%, 1%, 0.5%, 0.1%, according to the test kit in embodiment 1 and method to the solution D NA of this dilution for carrying out the detection sensitivity analysis.The pcr amplification condition provided according to embodiment 1 is tested, and result as shown in Figure 4.Experimental result shows, analyzes to detect to be limited to 5ng.
Claims (6)
1. an amplimer that detects the mink derived component, is characterized in that, its nucleotides sequence is classified as:
Forward primer: 5 '-CTTCAACCTCAACATCATCACC-3 ';
Reverse primer: 5 '-GACATACATTGTATTCATTCTAAGCG-3 '.
2. the amplimer of detection mink derived component as claimed in claim 1, is characterized in that, while with the mink genome, carrying out pcr amplification reaction, the nucleotide sequence of its amplified production is as shown in SEQ ID No.1.
3. the test kit that contains the amplimer of detection mink derived component claimed in claim 1.
4. a method that detects the mink derived component, it adopts amplimer claimed in claim 1 to carry out pcr amplification to mink DNA sample, and detects amplified production.
5. the method for detection mink derived component as claimed in claim 4, is characterized in that, the sample of the nucleotide sequence of amplified production as shown in SEQ ID No.1 is the mink sample.
6. the method for as described as claim 4 or 5 detection mink derived component, it is characterized in that: the PCR response procedures is: denaturation: 94 ℃, 3min; Enter circulation: 94 ℃ of sex change 60s, 66 ℃ of annealing 60s, 72 ℃ are extended 60s, 35 circulations; Stop extending: 72 ℃, 7min; Preserve reaction product for 4 ℃.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104032008B (en) * | 2014-06-12 | 2016-04-13 | 中国动物疫病预防控制中心 | The PCR primer pair of qualification or assistant identification recoon dog tissue and/or organ and application thereof |
| CN105018611B (en) * | 2015-07-14 | 2018-08-14 | 山东省兽药质量检验所 | A kind of LAMP detection method differentiating ermine meat in beef and mutton using mitochondrial DNA |
| CN107488705A (en) * | 2016-06-12 | 2017-12-19 | 中国检验检疫科学研究院 | The primed probe and method and kit precisely quantitatively detected for mink source composition digital pcr |
| CN107400717B (en) * | 2017-08-25 | 2020-10-27 | 广州迪澳医疗科技有限公司 | Constant-temperature fluorescence detection primer group, kit and detection method for mink-derived components |
| CN110117661A (en) * | 2018-02-07 | 2019-08-13 | 东北林业大学 | A kind of MGB Probe-detection methods that the sable fur products true and false identifies |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101434990A (en) * | 2008-12-15 | 2009-05-20 | 北华大学 | Mink heart DNA detection kit and identification method |
| CN101792816A (en) * | 2010-04-20 | 2010-08-04 | 东北林业大学 | Primer for amplifying marten species cytochrome b gene and method for identifying sable and pine marten |
| CN102337337A (en) * | 2011-09-28 | 2012-02-01 | 深圳出入境检验检疫局动植物检验检疫技术中心 | Polymerase chain reaction (PCR) detection primer for goose-derived component |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003204798A (en) * | 2001-10-31 | 2003-07-22 | Japan Spinners Inspecting Foundation | Method for discriminating animal hair fiber product |
-
2012
- 2012-04-11 CN CN201210105454.6A patent/CN102643912B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101434990A (en) * | 2008-12-15 | 2009-05-20 | 北华大学 | Mink heart DNA detection kit and identification method |
| CN101792816A (en) * | 2010-04-20 | 2010-08-04 | 东北林业大学 | Primer for amplifying marten species cytochrome b gene and method for identifying sable and pine marten |
| CN102337337A (en) * | 2011-09-28 | 2012-02-01 | 深圳出入境检验检疫局动植物检验检疫技术中心 | Polymerase chain reaction (PCR) detection primer for goose-derived component |
Non-Patent Citations (7)
| Title |
|---|
| JP特开2003-204798A 2003.07.22 |
| 张丽华等.貂组织线粒体DNA 及细胞色素b 鉴定及特征.《吉林大学学报》.2008,第34卷(第5期),第791页左栏1.1部分至第793页最后一段. |
| 杨明妍等.貂心与其伪品的脱氧核糖核酸指纹特征研究.《时珍国医国药》.2010,第21卷(第4期),937-938. |
| 水貂细胞色素 b 基因(Cytb)克隆、序 列分析及与鼬属动物的进化关系;涂剑锋等;《农业生物技术学报》;20101231;第18卷(第5期);975-980 * |
| 涂剑锋等.水貂细胞色素 b 基因(Cytb)克隆、序 列分析及与鼬属动物的进化关系.《农业生物技术学报》.2010,第18卷(第5期),975-980. |
| 貂心与其伪品的脱氧核糖核酸指纹特征研究;杨明妍等;《时珍国医国药》;20101231;第21卷(第4期);937-938 * |
| 貂组织线粒体DNA 及细胞色素b 鉴定及特征;张丽华等;《吉林大学学报》;20080930;第34卷(第5期);第791页左栏1.1部分至第793页最后一段 * |
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