CN109055573A - A method of based on the common reserve flour mite of multiple PCR technique Rapid identification - Google Patents
A method of based on the common reserve flour mite of multiple PCR technique Rapid identification Download PDFInfo
- Publication number
- CN109055573A CN109055573A CN201811081104.4A CN201811081104A CN109055573A CN 109055573 A CN109055573 A CN 109055573A CN 201811081104 A CN201811081104 A CN 201811081104A CN 109055573 A CN109055573 A CN 109055573A
- Authority
- CN
- China
- Prior art keywords
- mite
- pcr
- flour mite
- flour
- multiple pcr
- 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.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/16—Primer sets for multiplex assays
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a kind of methods based on the common reserve flour mite of multiple PCR technique Rapid identification, the genomic DNA of single flour mite is extracted first, using the DNA of extraction as pcr template, the specific primer of flour mite is designed on rDNA, under regular-PCR system and program, multiple PCR identification method is established, is detected by agarose gel electrophoresis, determines flour mite type；Show this method quickly, efficiently by various experimental verifications, specificity is high, compared with flour mite Morphological Identification method, is not necessarily to complete sample, and the children phases such as ovum, larva, nymph, hypopus, cryptic species, impaired sample can also identify；It is easy and economical compared with enzymatic cleavage methods and DNA sequencing method, a PCR reaction is only needed, the tedious steps of multiple PCR identification is avoided, alleviates workload, shorten detection time, general technical staff can operate.
Description
Technical field
The invention belongs to multiple PCR technique fields, and in particular to one kind is based on the common storage of multiple PCR technique Rapid identification
The method of object flour mite.
Background technique
Flour mite is under the jurisdiction of Acari (Acari), true mite catalogue (Acariformes), Astigmata
(Astigmata), individual is smaller, many kinds of, habitat is extensive, can largely multiply in reserve, mainly with plant or dynamic
Organic detritus of object is food, but based on grain storage, dry fruit, fur in reserve etc..Flour mite not only to reserve cause matter and
Very big loss in amount, but also Anaphylaxis disease can be caused, even result in pulmonary acariasis, intestines acariasis, urine acariasis etc..?
Under economic globalization, international trade and the frequent of tourism are even more provided convenience for the international communication of flour mite with diffusion with convenient,
The flour mite of known extensive occurring and damage has 98 kinds in world wide, wherein common flour mite has: Acarus siro (Acarus siro),
Tyrophagus putrescentiae (Tyrophagus putrescentiae), Na Shi elephant skin mite (Suidasia nesbitti), dermatophagoides pteronyssinus
(Dermatophagoidespteronyssinus), dust mite (Dermatophagoidesfarina) and Lepidogtyphusdestructor
(Lepidoglyphus destructor) etc..In recent years, flour mite is the harmful organism of intercepting and capturing of often being entered and left the border, and flour mite is quarantined
Through the importance for becoming grain storage Immigration and Quarantine.In the real-time detection of the grain storage flour mite of a large amount of inlet and outlet Agricultural Products Trades of entry and exit
In, the demand to a kind of simple, quick, economic, accurate identification technology is extremely urgent.
Currently, mainly being carried out according to its morphological feature in the identification of Immigration and Quarantine flour mite.Due to flour mite figure is small,
It is many kinds of, and such as children phase (including ovum, larva, nymph, hypopus) is frequently encountered, cryptic species and sample damage etc., quickly
Precise Identification is extremely difficult.In recent years, it is used widely based on molecular marking technique in species identification field.Wherein, with core
Spacer region 2 (ITS2) in the transcription of sugared body gene, because its evolutionary rate is very fast, even if its sequence length and alkali in nearly edge species
There is also larger differences for base composition, are widely applied as effective molecular labeling.Flour mite is carried out based on molecular marking technique
The method of identification has: polymerase chain reaction and PCR-RFLP, but above method complex steps, qualification time are long.
Summary of the invention
According to the above-mentioned deficiencies of the prior art, a kind of based on multiplex PCR the technical problem to be solved by the present invention is to propose
The method of the common reserve flour mite of technology Rapid identification, the present invention overcomes occur in the existing common reserve flour mite technology of identification
The problem of, the genomic DNA of single flour mite is extracted first, using the DNA of extraction as pcr template, carries out gram of ribosomes ITS sequence
Grand sequencing and analysis, the otherness of sequence length and base composition based on ITS2 devise 6 specific downstream primers, point
It is not named as Acas-R (Acarus siro), Tyrp-R (tyrophagus putrescentiae), Suin-R (Na Shi elephant skin mite), Derp-R (dermatophagoides pteronyssinus),
Derf-R (dust mite) and Lepd-R (Lepidogtyphusdestructor).Since 5.8S is very conservative between flour mite not of the same race, one is devised
General upstream primer of the Asti-F primer as 6 specific primers.On the basis of the reaction system of regular-PCR and program into
Row multiplexed PCR amplification realizes the identification to 6 kinds of common reserve flour mites simultaneously by agarose gel electrophoresis.And it is subsequent
It is demonstrated experimentally that the primer whether carries out regular-PCR specific amplification or carries out multiplex PCR specific amplification, the two amplification
As a result it is not significantly different, it was demonstrated that the multiple PCR primer specificity that the present invention designs is high.Only need a PCR reaction, so that it may
Amplify 6 kinds of different product lengths simultaneously, simply judge primer size can Direct Identification go out 6 kinds of flour mites.
In order to solve the above-mentioned technical problem, the technical solution adopted by the present invention are as follows:
A method of based on the common reserve flour mite of multiple PCR technique Rapid identification, comprising the following steps:
(1) genomic DNA of single flour mite is extracted；
(2) using the flour mite monomeric gene group DNA of extraction as template, ribosomes rDNA ITS sequence is expanded, based on the area ITS
Otherness site is designed primer of 7 primers mixing as PCR, is respectively as follows:
Asti-F:5 '-GTGGTGGATCACTCGGC-3 '；
Acas-R:5 '-GGCAAACATACATTGGCTCCC-3 '；
Tyrp-R:5 '-AGCCCCCTACATTAGACTACCA-3 '；
Suin-R:5 '-AGAGTCGACACGACCAACTG-3 '；
Derp-R:5 '-TGGAAATGACCTGACGACGTT-3 '；
Derf-R:5 '-TGGAAATGACCTGACGACGTT-3 '；
Lepd-R:5 '-ACCCGATCGATGCAATATGCT-3 '；
(3) on the basis of the reaction system of regular-PCR and program, 7 primers are added, carry out multiplexed PCR amplification reaction；
(4) to multi-PRC reaction product carry out electrophoresis detection, use 100+50bp DNALadder DNAMark J as
With reference to progress PCR product size judgement determines the type of flour mite: Acarus siro 429bp, tyrophagus putrescentiae 355bp, Na Shi
Elephant skin mite is 192bp, dermatophagoides pteronyssinus 215bp, dust mite 280bp, Lepidogtyphusdestructor 306bp.
Preferably, in 7 primers described in step (2), Asti-F is upstream primer, remaining 6 is under specificity
Swim primer.
Preferably, the condition of the reaction of regular-PCR described in step (3) and multiplexed PCR amplification reaction is equal are as follows: initial denaturation
94 DEG C, 3min；26 circulations: 94 DEG C, 30sec of denaturation is annealed 60 DEG C, 40sec, extends 72 DEG C, 1min；It terminates and extends 72 DEG C,
10min, finally in 4 DEG C of preservation 59min.
Preferably, electrophoresis described in step (4) is agarose gel electrophoresis, and the kind of flour mite is judged according to PCR product size
Class are as follows: Acarus siro 429bp, tyrophagus putrescentiae 355bp, Na Shi elephant skin mite are 192bp, dermatophagoides pteronyssinus 215bp, and dust mite is
280bp, Lepidogtyphusdestructor 306bp.
The medicine have the advantages that
1. the present invention disposably expands Acarus siro, tyrophagus putrescentiae, Na Shi elephant skin mite, room using multiple PCR technique simultaneously
Dust mite, dust mite, Lepidogtyphusdestructor specific gene, by agarose gel electrophoresis results it is simple judgement obtain identification knot
Fruit.
2. compared with traditional form identification, it is not necessarily to complete sample, residuum and children phase (ovum, larva and nymph) can also be with
Identification, qualification result accuracy and specificity are higher.Experimental result shows that, for same flour mite, which whether carries out
Regular-PCR specific amplification, or multiplex PCR specific amplification is carried out, the two result all has specificity, and amplification does not have
Significant difference.
3. it is easy and economical compared with enzymatic cleavage methods and DNA sequencing method, a PCR reaction is only needed, multiple PCR is avoided
The tedious steps of identification, alleviate workload, shorten detection time, and those skilled in the art can operate.
4. for the of less demanding of DNA profiling, even the DNA profiling of long-term preservation or template come from polypide fragment
There is good amplification.
5. experimental implementation is simple, reaction system is similar to program to regular-PCR, it is only necessary to a PCR reaction, so that it may same
When identify 6 kinds of common flour mites.Suitable for entry and exit port, accurate, the quick and high pass quantization of port quarantine is realized.
Detailed description of the invention
Content expressed by this specification attached drawing and the label in figure are briefly described below:
Fig. 1 is 6 kinds of flour mite PCR electrophoresis result schematic diagrames of the invention.
I: 6 kind of flour mite regular-PCR electrophoresis result, II: 6 kind of flour mite multiplex PCR electrophoresis result
1. Acarus siro, 2. tyrophagus putrescentiaes, 3. Na Shi elephant skin mites, 4. dermatophagoides pteronyssinus, 5. dust mites, 6. Lepidogtyphusdestructors
Specific embodiment
It below by the description to embodiment, is described in further detail, to help those skilled in the art to this hair
Bright inventive concept, technical solution have more complete, accurate and deep understanding.
Embodiment 1
6 kinds of flour mite monomeric gene group DNA are extracted using STE method respectively
(1) in the centrifuge tube containing single female mite, STE buffer (the 10mM Tris-HCl, 1mM of 20 μ L is added
EDTA, 100mM NaCl, PH=8.0), then respectively by centrifuge tube to grinding on ice；
(2) 0.5 μ L Proteinase K (25mg/mL) is added into centrifuge tube respectively, and mixes well；
(3) under room temperature, 1000g is centrifuged 1min, by mixed liquor in 56 DEG C of water-bath 60min；
(4) after the completion of water-bath, 95 DEG C of denaturation 5min, inactivated proteases K；
(5) under room temperature, 2000g is centrifuged 1min, the template for using the supernatant of 2 μ L to react as PCR, single flour mite base
After group DNA sample number, it is stored in spare in -20 DEG C of refrigerators.
Embodiment 2
The amplification of 2 groups of PCR is carried out, I group carries out regular-PCR amplification, II pair of 6 kinds of flour mite to 6 kinds of flour mite monomeric gene group DNA
Monomeric gene group carries out multiplexed PCR amplification.The two template is same monomer flour mite genomic DNA, a kind of flour mite corresponding one
A PCR reaction, reaction system volume is 25 μ L, and I group of specific reaction reagent and dosage are as shown in table 2；II group is specifically reacted examination
Agent and dosage are as shown in table 3:
26 kinds of flour mite regular-PCR amplification systems of table
36 kinds of flour mite multiplexed PCR amplification systems of table
6 kinds of flour mites are respectively Acarus siro, tyrophagus putrescentiae, Na Shi elephant skin mite, dermatophagoides pteronyssinus, dust mite and Lepidogtyphusdestructor；7 pairs
Primer is respectively Asti-F, Acas-R, Tyrp-R, Suin-R, Derp-R, Derf-R, Lepd-R；Response procedures are whether common
PCR or multiplex PCR are equal are as follows: 94 DEG C of 3min, 26 × (94 DEG C of 30sec, 60 DEG C of 40sec, 72 DEG C of 1min), 72 DEG C
10min, 4 DEG C of preservation 59min.
Embodiment 3
Multi-PRC reaction product is detected with 2% agarose gel electrophoresis, uses 100+50bp DNA
Ladder DNA Mark J carries out the judgement of PCR product size, determines the type of flour mite as reference.According to PCR product size
Judge the type of flour mite are as follows: Acarus siro 429bp, tyrophagus putrescentiae 355bp, Na Shi elephant skin mite are 192bp, and dermatophagoides pteronyssinus is
215bp, dust mite 280bp, Lepidogtyphusdestructor 306bp.
The present invention is exemplarily described above, it is clear that present invention specific implementation is not subject to the restrictions described above,
As long as using the improvement for the various unsubstantialities that the inventive concept and technical scheme of the present invention carry out, or not improved this is sent out
Bright conception and technical scheme directly apply to other occasions, within the scope of the present invention.Protection of the invention
Range should be determined by the scope of protection defined in the claims.
Claims (4)
1. a kind of method based on the common reserve flour mite of multiple PCR technique Rapid identification, which is characterized in that including following step
It is rapid:
(1) genomic DNA of single flour mite is extracted；
(2) using the flour mite monomeric gene group DNA of extraction as template, ribosomes rDNA ITS sequence, the difference based on the area ITS are expanded
Property site, design 7 primers and mix primer as PCR, be respectively as follows:
Asti-F:5 '-GTGGTGGATCACTCGGC-3 '；
Acas-R:5 '-GGCAAACATACATTGGCTCCC-3 '；
Tyrp-R:5 '-AGCCCCCTACATTAGACTACCA-3 '；
Suin-R:5 '-AGAGTCGACACGACCAACTG-3 '；
Derp-R:5 '-TGGAAATGACCTGACGACGTT-3 '；
Derf-R:5 '-TGGAAATGACCTGACGACGTT-3 '；
Lepd-R:5 '-ACCCGATCGATGCAATATGCT-3 '；
(3) on the basis of the reaction system of regular-PCR and program, 7 primers are added, carry out multiplexed PCR amplification reaction；
(4) electrophoresis detection is carried out to multi-PRC reaction product, uses 100+50bp DNALadder DNAMark J as ginseng
It examines, carries out the judgement of PCR product size, determine the type of flour mite.
2. a kind of method based on the common reserve flour mite of multiple PCR technique Rapid identification according to claim 1, special
Sign is that in 7 primers described in step (2), Asti-F is upstream primer, remaining 6 downstream primers for specificity.
3. a kind of method based on the common reserve flour mite of multiple PCR technique Rapid identification according to claim 1, special
Sign is that the condition of the reaction of regular-PCR described in step (3) and multiplexed PCR amplification reaction is equal are as follows: 94 DEG C of initial denaturation,
3min；26 circulations: 94 DEG C, 30sec of denaturation is annealed 60 DEG C, 40sec, extends 72 DEG C, 1min；It terminates and extends 72 DEG C, 10min,
Finally in 4 DEG C of preservation 59min.
4. a kind of method based on the common reserve flour mite of multiple PCR technique Rapid identification according to claim 1, special
Sign is that electrophoresis described in step (4) is agarose gel electrophoresis, the type of flour mite is judged according to PCR product size are as follows: thick foot
Flour mite is 429bp, and tyrophagus putrescentiae 355bp, Na Shi elephant skin mite is 192bp, dermatophagoides pteronyssinus 215bp, dust mite 280bp, evil
Thermophilic squama mite is 306bp.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811081104.4A CN109055573B (en) | 2018-09-17 | 2018-09-17 | Method for rapidly identifying common storage spider mites based on multiple PCR technology |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811081104.4A CN109055573B (en) | 2018-09-17 | 2018-09-17 | Method for rapidly identifying common storage spider mites based on multiple PCR technology |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109055573A true CN109055573A (en) | 2018-12-21 |
| CN109055573B CN109055573B (en) | 2021-12-14 |
Family
ID=64762714
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811081104.4A Active CN109055573B (en) | 2018-09-17 | 2018-09-17 | Method for rapidly identifying common storage spider mites based on multiple PCR technology |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109055573B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110117666A (en) * | 2019-06-06 | 2019-08-13 | 中国农业大学 | Identify the primer pair and its application of tyrophagus putrescentiae and ellipse dispersion |
| CN113817838A (en) * | 2021-08-31 | 2021-12-21 | 皖南医学院 | Dust mite microsatellite marker, primer and application thereof, and primer acquisition method |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1820006A (en) * | 2003-05-09 | 2006-08-16 | 拜尔农作物科学股份公司 | Substituted oxyarenes, and use thereof for controlling pests |
| CN103805708A (en) * | 2014-02-20 | 2014-05-21 | 南京农业大学 | PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) identification method for Chinese tetranychus common species |
| CN104521828A (en) * | 2014-12-18 | 2015-04-22 | 中国科学院水生生物研究所 | Method for building gyrodactylus purebred artificial infection system |
| US20160194725A1 (en) * | 2013-07-16 | 2016-07-07 | ALK-Ablló S/A | Molecular identification of allergy causing mites by pcr |
| ES2674128A1 (en) * | 2016-12-27 | 2018-06-27 | Universidad De Salamanca | Method for diagnosing allergic sensitization in a subject (Machine-translation by Google Translate, not legally binding) |
-
2018
- 2018-09-17 CN CN201811081104.4A patent/CN109055573B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1820006A (en) * | 2003-05-09 | 2006-08-16 | 拜尔农作物科学股份公司 | Substituted oxyarenes, and use thereof for controlling pests |
| US20160194725A1 (en) * | 2013-07-16 | 2016-07-07 | ALK-Ablló S/A | Molecular identification of allergy causing mites by pcr |
| CN103805708A (en) * | 2014-02-20 | 2014-05-21 | 南京农业大学 | PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) identification method for Chinese tetranychus common species |
| CN104521828A (en) * | 2014-12-18 | 2015-04-22 | 中国科学院水生生物研究所 | Method for building gyrodactylus purebred artificial infection system |
| ES2674128A1 (en) * | 2016-12-27 | 2018-06-27 | Universidad De Salamanca | Method for diagnosing allergic sensitization in a subject (Machine-translation by Google Translate, not legally binding) |
Non-Patent Citations (4)
| Title |
|---|
| B.BEROIZ等: ""Mite species identification in the production of allergenic extracts for clinical use and in environmental samples by ribosomal DNA amplification"", 《MEDICAL AND VETERINARY ENTOMOLOGY》 * |
| 程训佳等: "《人体寄生虫学》", 30 September 2015, 复旦大学出版社 * |
| 郑凌霄: ""常见储藏物粉螨的分子鉴定与系统发育研究"", 《中国优秀硕士学位论文全文数据库基础科学辑》 * |
| 郑凌霄等: ""基于DNA条形码技术的粉螨种类鉴定研究"", 《中国媒介生物学及控制杂志》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110117666A (en) * | 2019-06-06 | 2019-08-13 | 中国农业大学 | Identify the primer pair and its application of tyrophagus putrescentiae and ellipse dispersion |
| CN110117666B (en) * | 2019-06-06 | 2022-04-26 | 中国农业大学 | Specific primer pair for identifying Tyrophagus putrescentiae and Tyrophagus ovatus and application thereof |
| CN113817838A (en) * | 2021-08-31 | 2021-12-21 | 皖南医学院 | Dust mite microsatellite marker, primer and application thereof, and primer acquisition method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109055573B (en) | 2021-12-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101696452A (en) | Method for identifying molecules of recessive white feather genes of chicken | |
| Zhao et al. | Molecular genetic diversity and population structure in Lycium accessions using SSR markers | |
| CN105177148B (en) | The double PCR primer of two kinds of grape ulcer bacterium of detection and its application simultaneously | |
| CN109055573A (en) | A method of based on the common reserve flour mite of multiple PCR technique Rapid identification | |
| CN112695125B (en) | Katelia SSR molecular marker primer composition and application thereof | |
| CN112695124B (en) | Phalaenopsis SSR molecular marker primer composition and application thereof | |
| CN108950088A (en) | A kind of double PCR primer and method identifying grapevine leafroll virus 3 and grape Stem Pitting Virus | |
| CN112195262A (en) | ISSR-PCR (inter-simple sequence repeat-polymerase chain reaction) amplification reaction system of Hubei fritillary bulb, ISSR-PCR molecular marking method and application | |
| CN106701968A (en) | Primer for simultaneously detecting Botryosphaeria dothidea and Cryptosporella viticola and application of primer | |
| Grube et al. | Siphula represents a remarkable case of morphological convergence in sterile lichens | |
| CN107326082B (en) | Universal primer of human cestode ep45 gene and identification method thereof | |
| CN104120170B (en) | Potato early blight bacterium detection kit and detection method | |
| CN106755526B (en) | Functional molecular marker related to lipped and petal sepal character of cymbidium and identification method thereof | |
| CN102242225B (en) | Double RT-PCR (reverse transcription-polymerase chain reaction) detection method of CVB (chrysanthemum virus B) and CChMVd (chrysanthemum chlorotic mottle viroid) | |
| CN113403417B (en) | SSR molecular marker AerM01 for sex identification of actinidia arguta and application thereof | |
| CN108841983A (en) | A kind of SSR primer of sugarcane overall length transcript profile data large-scale development | |
| CN114807411A (en) | Method for detecting sex of kiwi fruits | |
| CN107523644B (en) | Identification method of bletilla ochracea | |
| CN112680542A (en) | Universal SSR molecular marker primer composition for orchidaceae plants and application of universal SSR molecular marker primer composition | |
| CN102181563B (en) | Method for identifying common warehousing liposcelis quickly based on multiple PCR technology | |
| CN104946742A (en) | SPAR (single primer amplification reaction) based DNA molecular marking method | |
| CN110273023B (en) | Method for identifying or distinguishing cinnamon and related plants thereof, and primers and kit used by method | |
| CN112410454B (en) | Primer and method for identifying rhododendron dauricum and rhododendron lapponicum | |
| KR102081586B1 (en) | Housekeeping gene of onion, and method for separating the same | |
| CN112899393A (en) | Codonopsis species cpPSSR molecular marker and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |