CN101434995A - Northeast wood frog and molecular identification method for egg oil thereof - Google Patents
Northeast wood frog and molecular identification method for egg oil thereof Download PDFInfo
- Publication number
- CN101434995A CN101434995A CNA2008102296207A CN200810229620A CN101434995A CN 101434995 A CN101434995 A CN 101434995A CN A2008102296207 A CNA2008102296207 A CN A2008102296207A CN 200810229620 A CN200810229620 A CN 200810229620A CN 101434995 A CN101434995 A CN 101434995A
- Authority
- CN
- China
- Prior art keywords
- frog
- dna
- minutes
- add
- ovum oil
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
A molecular identification method for the wood frog in the northeast of China and the ovum oil thereof aims at solving the identification of the wood frog in the northeast of China and the product ovum oil thereof, which is mainly based on the property characteristics, microscopic characteristics, physical and chemical properties, and the like at present; as the bases are lack of clear shape identification characteristics and special chemical compositions, the identification for the ovum oil in practice is still very difficult and the invention is hereby designed. The identification method takes PCR technology as the basis and screens specific primers for amplification so as to achieve the molecular identification of the wood frog in the northeast of China and the ovum oil thereof, comprising the following steps: (1) extraction of genomic DNA; (2) genomic DNA detection and concentration determination; (3) PCR amplification; and (4) PCR amplification product identification. The invention is characterized in that the invention starts from the basic theory of molecular biology, uses modern molecule marker technology for conducting molecular identification of the wood frog in the northeast of China and the ovum oil thereof, has the advantages of scientific and credible theoretical basis, advanced and reasonable technical means and achieving steps, reliable detection result, strong specificity of screened primers as well as high stability of agarose gel electrophoresis characteristic bands, and can obtain reliable and useful effects in practical application.
Description
Technical field:
The present invention relates to a kind of molecular identification method of biology, be primarily aimed at the molecular identification method of Dongbei forest-frog animal kind and products thereof ovum oil.
Background technology:
Dongbei forest-frog has important economic implications and value medical health care.The fallopian tube of female frog dry products is used as medicine, and is called wood frog oil or Oviductus Ranae, is famous and precious animal drug.This medicinal material is identified at present still mainly according to characteristic trait, microscopic features, physico-chemical property etc., but because this medicinal material lacks tangible form diagnostic characteristics and special chemical ingredients, in the practice wood frog ovum oil identifying is still felt very difficult.
Summary of the invention:
In the production and operation practice, be difficult to the problem identified from aspects such as characteristic trait, microscopic features, physico-chemical properties at Dongbei forest-frog and woodfrog egg oil, applied molecular biology technology of the present invention solves the Molecular Identification of woodfrog egg oil and medicine source animal Dongbei forest-frog thereof, provide a kind of based on round pcr, the screening special primer increases, thereby reach the molecular identification method to Dongbei forest-frog and ovum oil thereof, the specific implementation step is:
(1) extracting genome DNA:
1. take by weighing tissue sample 100～500mg, shred, put into the EP pipe of the 1.5ml of sterilization;
2. add 600 μ l, 1 * SET liquid, 20 μ l Proteinase Ks (10mg/ml), 25 μ l SDS, temperature under 55 ℃ condition, airbath 2 hours;
3. add the 500 saturated phenol of μ l Tris and 200 μ l TE turned upside down 10 minutes;
4. with 10000 rev/mins, centrifugal 10 minutes, suct a layer water, lower floor removes;
5. add isopyknic phenol: chloroform: primary isoamyl alcohol (24:23:1), add 200 μ l TE again and mixed 10 minutes;
6. with 10000 rev/mins, centrifugal 10 minutes, suct a layer water, lower floor removes;
7. add isopyknic chloroform: primary isoamyl alcohol (23:1), add 200 μ l TE again and mixed 10 minutes;
8. with 10000 rev/mins, centrifugal 10 minutes, suct a layer water, lower floor removes;
9. add the dehydrated alcohol (20 ℃ of precoolings) of 2 times of volumes, level is shaken 30 times, goes out white DNA precipitation with the rifle choicest;
10. the cold ethanol that adds 700 μ l 70% with 8000 rev/mins, centrifugal 5 minutes, is outwelled ethanol, seasoning;
Add TE solution 50～100 μ l, place 55 ℃ of water-baths 3～4 hours;
(2) genomic dna detects and concentration determination
Measure DNA concentration with ultraviolet spectrophotometer: according to the OD260/280 ratio of record, estimation DNA quality, OD260/280 can satisfy test requirements document between 1.6～1.8; According to the OD260 numerical value and the test fluid extension rate of record, converse the concentration of DNA.DNA stoste is done corresponding dilution,, preserve use for 4 ℃ as working fluid with 50ng/ μ l packing.
(3) pcr amplification
The employing special primer (F:5 '-GCC TTT CTA CCG CAA ATA-3 '; R:5 '-AGG TGCTTT TTT TCT GGG-3 ') in the PCR instrument, increases.
(4) the specific amplified product is differentiated
Amplified production detects through 1.2% agarose gel electrophoresis, 100V constant voltage, EB dyeing; On the gel pattern analytical system, observe and analyze, and the record result; Dongbei forest-frog and ovum oil genomic dna amplify specific band at the 1000bp place, and asexuality and individual difference are as the key band of Dongbei forest-frog and the discriminating of ovum oil.
Characteristics of the present invention and beneficial effect:
The present invention is from the molecular biology basic theories, uses modern molecular marking technique and Dongbei forest-frog and ovum oil are carried out molecule differentiates that it is credible to have the theoretical basis science, and technique means and performing step are rationally advanced, and characteristics such as detected result is reliable.Among the present invention, the detection of the screening of pcr amplification primer and agarose gel electrophoresis key band is a key link, repeatedly detect evidence repeatedly, the primer specificity of screening strong, agarose gel electrophoresis key band (1000bp) stability is high, can obtain reliable and useful effect in actual applications.
Description of drawings:
Fig. 1 wood frog sample special primer DNA cloning collection of illustrative plates
Embodiment:
The molecular identification method of Dongbei forest-frog and ovum oil thereof, this method are a kind of based on round pcr, and the screening special primer increases, and realize the molecule of Dongbei forest-frog and ovum oil is differentiated that the specific implementation step is:
(1) extracting genome DNA:
1. take by weighing tissue sample 100～500mg, shred, put into the EP pipe of the 1.5ml of sterilization;
2. add 600 μ l, 1 * SET liquid, 20 μ l Proteinase Ks (10mg/ml), 25 μ l SDS, temperature under 55 ℃ condition, airbath 2 hours;
3. add the 500 saturated phenol of μ l Tris and 200 μ l TE turned upside down 10 minutes;
4. with 10000 rev/mins, centrifugal 10 minutes, suct a layer water, lower floor removes;
5. add isopyknic phenol: chloroform: primary isoamyl alcohol (24:23:1), add 200 μ l TE again and mixed 10 minutes;
6. with 10000 rev/mins, centrifugal 10 minutes, suct a layer water, lower floor removes;
7. add isopyknic chloroform: primary isoamyl alcohol (23:1), add 200 μ l TE again and mixed 10 minutes;
8. with 10000 rev/mins, centrifugal 10 minutes, inhale upper water, phase lower floor removes;
9. add the dehydrated alcohol (20 ℃ of precoolings) of 2 times of volumes, level is shaken 30 times, goes out white DNA precipitation with the rifle choicest;
10. the cold ethanol that adds 700 μ l 70% with 8000 rev/mins, centrifugal 5 minutes, is outwelled ethanol, seasoning;
Preserve down at 4 ℃.
(2) genomic dna detects and concentration determination
Measure DNA concentration with ultraviolet spectrophotometer: according to the OD260/280 ratio of record, estimation DNA quality, general OD260/280 can satisfy test requirements document between 1.6～1.8; According to the OD260 numerical value and the test fluid extension rate of record, converse the concentration of DNA.DNA stoste is done corresponding dilution, with 50ng/ μ l packing, preserve use for 4 ℃ as working fluid, DNA stoste places-20 ℃ of preservations.
(3) pcr amplification
With primer (F:5 '-GCC TTT CTA CCG CAA ATA-3 '; R:5 '-AGG TGC TTT TTTTCT GGG-3 ') in the PCR instrument, carries out the specific amplified reaction;
The pcr amplification reaction system is formed:
10 * PCR buffer is 2.5 μ l, 10mmol/L dNTP is 2.0 μ l, Primer (F) 10pmol/ μ l is 1.0 μ l, Primer (R) 10pmol/ μ l is 1.0 μ l, Taq DNA polymerase is 0.2 μ l (1U), Template is 2.0 μ l (50ng/ μ l), and Autoclaved, distilled water are 16.3 μ l.Cumulative volume is 25ul.
The pcr amplification reaction program:
At 94.0 ℃ of pre-sex change 7min, enter following circulation then: 94.0 ℃ of sex change 50s, 56.9 ℃ of renaturation 1min, 72.0 ℃ of extension 1min, circulate 35 times, 72.0 ℃ are extended 10min again after the loop ends.(4) the specific amplified product is differentiated
Amplified production detects through 1.2% agarose gel electrophoresis, 100V constant voltage, EB dyeing; On the gel pattern analytical system, observe and analyze, and the record result; Dongbei forest-frog and ovum oil genomic dna amplify specific band at the 1000bp place, and asexuality and individual difference can be used as the key band that Dongbei forest-frog and ovum oil is differentiated.
Embodiment:
Buying is made to differentiate object from the geographic Dongbei forest-frog of Changbai mountain, Jilin and the ovum oil true and false.
Dongbei forest-frog and ovum oil sample are originally respectively got 3.The concrete operations step is as follows:
(1) extracting genome DNA:
1. use electronic balance (FA1604N, Shanghai exact science instrument plant) take by weighing sample and organize 200mg (the Dongbei forest-frog sample is got muscle tissue), use 70% alcohol wash, the sterilization scissors shreds, put into the EP pipe of 1.5ml, natural air drying made the alcohol volatilization in 30～60 minutes;
2. in the EP pipe, add 600 μ l, 1 * SET liquid, 20 μ l Proteinase Ks (10mg/ml), 25 μ 1SDS, with 55 ℃ of following airbaths of airbath vibrator (CHA-S, all over the country company limited in Shenzhen) 2 hours;
3. adding 500 saturated phenol of μ l Tris and 200 μ l TE turned upside down 10 minutes in the EP pipe;
4. use supercentrifuge (Biofuge primo R, German SORVALL company) with 10000 rev/mins, centrifugal 10 minutes, draw the upper strata water, lower floor removes;
5. add isopyknic phenol: chloroform: primary isoamyl alcohol (24:23:1), add 200 μ l TE liquid again and mixed 10 minutes;
6. use supercentrifuge with 10000 rev/mins, centrifugal 10 minutes, draw the upper strata water, lower floor removes;
7. add isopyknic chloroform: primary isoamyl alcohol (23:1), add 200 μ l TE liquid again and mixed 10 minutes;
8. use supercentrifuge with 10000 rev/mins, centrifugal 10 minutes, suct a layer water, lower floor removes;
9. add the dehydrated alcohol (20 ℃ of precoolings) of 2 times of volumes, level is shaken 30 times, goes out white DNA precipitation with the rifle choicest;
10. the cold ethanol that adds 700 μ l 70% with 8000 rev/mins, centrifugal 5 minutes, is outwelled ethanol, seasoning with supercentrifuge;
Preserve down at 4 ℃.
(2) genomic dna detects and concentration determination
Measure DNA concentration with ultraviolet spectrophotometer (OPTIZEN 2120UV, Korea S Mei Kaxisi): the OD260/280 ratio of mensuration illustrates that the DNA quality is better between 1.70～1.80, can satisfy test requirements document; Going out DNA concentration according to the numerical evaluation of OD260 is 120ng/ μ l.Be diluted to 50ng/ μ l packing, preserve for 4 ℃ as working fluid and use.
(3) pcr amplification
With primer (F:5 '-GCC TTT CTA CCG CAA ATA-3 '; R:5 '-AGG TGC TTT TTTTCT GGG-3 ') (the big Gene science company of Beijing six directions China is synthetic) is in PCR instrument (MyCycler
TMThermalCycler carries out the specific amplified reaction in Bio-rad);
The pcr amplification reaction system is formed:
10 * PCR buffer is 2.5 μ l, 10mmol/L dNTP is 2.0 μ l, Primer (F) 10pmol/ μ l is 1.0 μ l, Primer (R) 10pmol/ μ l is 1.0 μ l, Taq DNA polymerase is 0.2 μ l (1U), Template is 2.0 μ l (50ng/ μ l), sterilization distilled water (self-control of Mylli-Q Biocel ultrapure water instrument) 16.3 μ l.Cumulative volume is 25ul.PCR buffer, dNTP and Taq DNApolymerase are available from Generay Biotech (Shanghai) Co.Ltd.
The pcr amplification reaction program:
At 94.0 ℃ of pre-sex change 7min, enter following circulation then: 94.0 ℃ of sex change 50s, 56.9 ℃ of renaturation 1min, 72.0 ℃ of extension 1min, circulate 35 times, 72.0 ℃ are extended 10min again after the loop ends.
(4) the specific amplified product is differentiated
Amplified production EC250-90 (U.S. power ﹠ light company) electrophoresis apparatus electrophoresis detection.Sepharose concentration 1.2%, add the pcr amplification product and the dna molecular amount mark (DNA Marker DL2000 is available from Shanghai Generay Biotech Co.Ltd) of 6 samples in the well respectively, set a negative control simultaneously, 100V constant voltage electrophoresis, EB dyeing.Referring to Fig. 1, at (the 440CF of gel imaging analysis system, Kodak) go up observations, these 5,6 genomic dnas of wood frog sample 1,2 and ovum oil sample amplify clear band at 1000bp place, can assert that institute's sample that detects really is the oily genuine piece of Dongbei forest-frog and ovum; Wood frog sample 3 and ovum oil sample basis 4 assert that then at the no key band in 1000bp place the sample that detects is Dongbei forest-frog and the pseudo-product of ovum oil.7 negative contrasts among Fig. 1, M is DNA Marker.
Claims (3)
1, the molecular identification method of Dongbei forest-frog and ovum oil thereof is characterized in that: this method is a kind of based on round pcr, and the screening special primer increases, and realizes the molecule of Dongbei forest-frog and ovum oil is differentiated that the specific implementation step is:
(1) extracting genome DNA:
1. take by weighing tissue sample 100～500mg, shred, put into the EP pipe of the 1.5ml of sterilization;
2. add 600 μ l, 1 * SET liquid, concentration is Proteinase K 20 μ l, the 25 μ l SDS of 10mg/ml, temperature under 55 ℃ condition, airbath 2 hours;
3. add the 500 saturated phenol of μ l Tris and 200 μ l TE turned upside down 10 minutes;
4. with 10000 rev/mins, centrifugal 10 minutes, suct a layer water, lower floor removes;
5. add isopyknic phenol: chloroform: primary isoamyl alcohol=24:23:1, add 200 μ l TE again and mixed 10 minutes;
6. with 10000 rev/mins, centrifugal 10 minutes, suct a layer water, lower floor removes;
7. add isopyknic chloroform: primary isoamyl alcohol=23:1, add 200 μ l TE again and mixed 10 minutes;
8. with 10000 rev/mins, centrifugal 10 minutes, suct a layer water, lower floor removes;
9. precooling under-20 ℃ of temperature, the dehydrated alcohol of 2 times of volumes of adding, level is shaken 30 times, goes out white DNA precipitation with the rifle choicest;
10. the cold ethanol that adds 700 μ l 70% with 8000 rev/mins, centrifugal 5 minutes, is outwelled ethanol, seasoning;
(2) genomic dna detects and concentration determination
Measure DNA concentration with ultraviolet spectrophotometer: according to the OD260/280 ratio of record, estimation DNA quality, OD260/280 can satisfy test requirements document between 1.6～1.8; According to the OD260 numerical value and the test fluid extension rate of record, converse the concentration of DNA.DNA stoste is done corresponding dilution,, preserve use for 4 ℃ as working fluid with 50ng/ μ l packing.
(3) pcr amplification
The employing special primer (F:5 '-GCC TTT CTA CCG CAA ATA-3 '; R:5 '-AGG TGCTTT TTT TCT GGG-3 ') in the PCR instrument, increases.
(4) the specific amplified product is differentiated
Amplified production detects through 1.2% agarose gel electrophoresis, 100V constant voltage, EB dyeing; On the gel pattern analytical system, observe and analyze, and the record result; Dongbei forest-frog and ovum oil genomic dna amplify specific band at the 1000bp place, and asexuality and individual difference are as the key band of Dongbei forest-frog and the discriminating of ovum oil.
2, the molecular identification method of Dongbei forest-frog according to claim 1 and ovum oil thereof, it is characterized in that: described pcr amplification reaction cumulative volume is 25ul, comprising following each component: 10 * PCR buffer is 2.5 μ l, 10mmol/L dNTP is 2.0 μ l, Primer (F) 10pmol/ μ l is 1.0 μ l, Primer (R) 10pmol/ μ l is 1.0 μ l, Taq DNA polymerase is 0.2 μ l (1U), Template is 2.0 μ l (50ng/ μ l), Autoclaved, distilled water are 16.3 μ l.
3, the molecular identification method of Dongbei forest-frog according to claim 1 and ovum oil thereof, it is characterized in that: described pcr amplification reaction program is, at 94.0 ℃ of pre-sex change 7min, enter following circulation then: 94.0 ℃ of sex change 50s, 56.9 ℃ of renaturation 1min, 72.0 ℃ of extension 1min, circulate 35 times, 72.0 ℃ are extended 10min again after the loop ends.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008102296207A CN101434995A (en) | 2008-12-11 | 2008-12-11 | Northeast wood frog and molecular identification method for egg oil thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008102296207A CN101434995A (en) | 2008-12-11 | 2008-12-11 | Northeast wood frog and molecular identification method for egg oil thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101434995A true CN101434995A (en) | 2009-05-20 |
Family
ID=40709631
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2008102296207A Pending CN101434995A (en) | 2008-12-11 | 2008-12-11 | Northeast wood frog and molecular identification method for egg oil thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101434995A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106350510A (en) * | 2016-10-17 | 2017-01-25 | 哈尔滨市食品药品检验检测中心 | Method and kit for quickly extracting genome DNA from forest frog's oviduct, and application thereof |
| CN106636399A (en) * | 2016-12-21 | 2017-05-10 | 中国中医科学院中药研究所 | Method for performing true and false identification on oviductus ranae and special primer |
| CN106755452A (en) * | 2017-01-05 | 2017-05-31 | 中国医学科学院药用植物研究所 | Oviductus Ranae medicinal material DNA bar code identifies pcr amplification primer thing and amplification method |
| CN110964798A (en) * | 2020-03-05 | 2020-04-07 | 东北农业大学 | Method for identifying northeast wood frog genetic sex by using TRAP molecular marker technology |
| CN111593128A (en) * | 2020-04-13 | 2020-08-28 | 东北农业大学 | Molecular marker for identifying northeast wood frog genetic sex and application method thereof |
| CN111690658A (en) * | 2020-05-26 | 2020-09-22 | 长春海关技术中心 | DNA extraction and identification method of rana japonica oil |
| CN112725458A (en) * | 2019-10-28 | 2021-04-30 | 成都市食品药品检验研究院 | PCR kit and PCR method for identifying oviductus ranae |
-
2008
- 2008-12-11 CN CNA2008102296207A patent/CN101434995A/en active Pending
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106350510B (en) * | 2016-10-17 | 2019-05-10 | 哈尔滨市食品药品检验检测中心 | Method, kit and its application of rapidly extracting Oviductus Ranae genomic DNA |
| CN106350510A (en) * | 2016-10-17 | 2017-01-25 | 哈尔滨市食品药品检验检测中心 | Method and kit for quickly extracting genome DNA from forest frog's oviduct, and application thereof |
| CN106636399A (en) * | 2016-12-21 | 2017-05-10 | 中国中医科学院中药研究所 | Method for performing true and false identification on oviductus ranae and special primer |
| CN106636399B (en) * | 2016-12-21 | 2020-03-03 | 中国中医科学院中药研究所 | Method for identifying authenticity of oviductus ranae and special primer |
| CN106755452B (en) * | 2017-01-05 | 2021-04-02 | 中国医学科学院药用植物研究所 | PCR amplification primer and amplification method for identifying oviductus ranae medicinal material DNA bar code |
| CN106755452A (en) * | 2017-01-05 | 2017-05-31 | 中国医学科学院药用植物研究所 | Oviductus Ranae medicinal material DNA bar code identifies pcr amplification primer thing and amplification method |
| CN112725458A (en) * | 2019-10-28 | 2021-04-30 | 成都市食品药品检验研究院 | PCR kit and PCR method for identifying oviductus ranae |
| CN110964798A (en) * | 2020-03-05 | 2020-04-07 | 东北农业大学 | Method for identifying northeast wood frog genetic sex by using TRAP molecular marker technology |
| CN110964798B (en) * | 2020-03-05 | 2023-07-07 | 东北农业大学 | Method for identifying genetic sex of rana chensinensis by using TRAP molecular marker technology |
| CN111593128A (en) * | 2020-04-13 | 2020-08-28 | 东北农业大学 | Molecular marker for identifying northeast wood frog genetic sex and application method thereof |
| CN111593128B (en) * | 2020-04-13 | 2024-06-11 | 东北农业大学 | Molecular marker for identifying genetic sex of northeast wood frog and application method thereof |
| CN111690658A (en) * | 2020-05-26 | 2020-09-22 | 长春海关技术中心 | DNA extraction and identification method of rana japonica oil |
| CN111690658B (en) * | 2020-05-26 | 2023-12-22 | 长春海关技术中心 | DNA extraction and identification method of oviductus ranae |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101434995A (en) | Northeast wood frog and molecular identification method for egg oil thereof | |
| KR20130045636A (en) | Ssr primer derived from paeonia lactiflora and use thereof | |
| CN103898195A (en) | Helicobacter pylori drug resistance nucleic acid detection kit | |
| CN104059975B (en) | To Providence O3, the Nucleotide that O4, O8, O12, O13 and O20 are special and application thereof | |
| Harasawa et al. | Molecular evidence for hemotropic Mycoplasma infection in a Japanese badger (Meles meles anakuma) and a raccoon dog (Nyctereutes procyonoides viverrinus) | |
| CN111850134A (en) | Specific forward and reverse primers and probe for rainbow trout, detection kit and application of specific forward and reverse primers and probe | |
| CN106282373A (en) | A kind of Herba Dendrobii and the contrast authentication method of Henan Herba Dendrobii | |
| CN107988422A (en) | With soya seeds oil content relevant SNP marker, section, primer and application | |
| Sun et al. | Database and primer selections affect nematode community composition under different vegetations of Changbai Mountain | |
| CN107400736A (en) | The type adenovirus ring mediated isothermal amplification detection primer group of duck 2 and kit | |
| Abd-Elsalam et al. | Genetic characterization of Fusarium oxysporum f. sp. vasinfectum isolates by random amplification of polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP)/Genetische Charakterisierung von Fusarium oxysporum f. sp. vasinfectum-Isolaten durch RAPD (random amplification of polymorphic DNA) und AFLP (amplified fragment length polymorphism) | |
| Jiang et al. | Sap-direct RT-PCR for the rapid detection of coleus blumei viroids of the genus Coleviroid from natural host plants | |
| Sun et al. | Molecular typing of epizootic hemorrhagic disease virus serotypes by one-step multiplex RT-PCR | |
| CN108866205A (en) | Identify the specific primer of hiruto based on molecular biology method | |
| CN103146834A (en) | Allglo probe-based detection method of anopheles sinensis knockdown resistance gene mutation site | |
| CN101712983A (en) | Primer pair for detecting citrus canker pathogenic bacteria and detection method thereof | |
| CN109486983A (en) | Buta-buta, Yunnan agalloch eaglewood, Aguilaria malaccensis Lamk SNP marker and application | |
| CN102719543B (en) | Method for identifying plant varieties by utilizing chemical molecular formulas of nucleotides | |
| CN105132562B (en) | For identifying molecular labeling, primer pair and its application of the non-acid character of Peach fruits | |
| CN103667539A (en) | Kit for detecting peste des petits ruminants virus by utilizing pyrophosphoric-acid sequencing technology | |
| CN103993090A (en) | Specific nucleotides for providencia O31, O41, O42, O43 and O50 and application of specific nucleotides | |
| CN104073560B (en) | The rapid molecular discrimination method of a kind of Eucommia ulmoides or former plant | |
| CN103290107A (en) | Construction method of SSR fingerprint suitable for identification of rice hybrid authenticity | |
| CN103923981B (en) | HLA-B*27 Allele Detection Method and kit thereof | |
| KR101695051B1 (en) | Universal primer set COS0842 for discrimination of Brassicaceae sp. and molecular marker comprising the same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20090520 |