CN105543364B - A kind of specific gene and its method for identifying molecules of Thailand's Storehouse midge - Google Patents
A kind of specific gene and its method for identifying molecules of Thailand's Storehouse midge Download PDFInfo
- Publication number
- CN105543364B CN105543364B CN201610012014.4A CN201610012014A CN105543364B CN 105543364 B CN105543364 B CN 105543364B CN 201610012014 A CN201610012014 A CN 201610012014A CN 105543364 B CN105543364 B CN 105543364B
- Authority
- CN
- China
- Prior art keywords
- thailand
- storehouse midge
- gene
- midge
- rdna
- 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.)
- Expired - Fee Related
Links
- 241000256135 Chironomus thummi Species 0.000 title claims abstract description 57
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 43
- 238000000034 method Methods 0.000 title claims abstract description 26
- 108020004465 16S ribosomal RNA Proteins 0.000 claims abstract description 22
- 238000012408 PCR amplification Methods 0.000 claims description 13
- 241000238631 Hexapoda Species 0.000 claims description 6
- 238000001962 electrophoresis Methods 0.000 claims description 6
- 238000012163 sequencing technique Methods 0.000 claims description 5
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 claims description 4
- 229960005542 ethidium bromide Drugs 0.000 claims description 4
- 238000000605 extraction Methods 0.000 claims description 4
- 238000010186 staining Methods 0.000 claims description 3
- 229920000936 Agarose Polymers 0.000 claims description 2
- 230000003321 amplification Effects 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- 210000002966 serum Anatomy 0.000 claims description 2
- 238000012360 testing method Methods 0.000 claims description 2
- 241000894007 species Species 0.000 abstract description 7
- 108020004414 DNA Proteins 0.000 description 15
- 239000008280 blood Substances 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 230000000877 morphologic effect Effects 0.000 description 6
- 230000009182 swimming Effects 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 238000000246 agarose gel electrophoresis Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 238000012797 qualification Methods 0.000 description 3
- 241000134316 Culicoides <genus> Species 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 2
- 108020005196 Mitochondrial DNA Proteins 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 239000012154 double-distilled water Substances 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 238000010170 biological method Methods 0.000 description 1
- 208000003836 bluetongue Diseases 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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
- 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- 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
- C12Q2600/158—Expression markers
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (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)
Abstract
The present invention discloses a kind of specific gene and its method for identifying molecules of Thailand's Storehouse midge, it is characterised in that the 16S rDNA gene orders that the Storehouse midge is obtained using molecular biology method carry out Identification of Species by the comparison of gene order similitude to the Storehouse midge.Thailand's Storehouse midge method for identifying molecules provided by the invention is advantageously implemented Thailand's Storehouse midge and accurately and rapidly identifies.
Description
Technical field
The present invention relates to species identification fields, and in particular to the special gene sequence of Thailand's Storehouse midge and its Molecular Identification side
Method.
Background technology
Storehouse midge is one of three big blood sucking midge of China, can propagate blue tongue disease, live in a variety of Amphixenosis such as globidiosis, is a kind of
Important Vector.Therefore, to the identification of blood sucking midge and identification, it is that monitoring passes the pathogenetic basis of disease with control midge, and
The core procedure in prevention and control of diseases field.Know currently, relying primarily on veteran taxology expert to the identification of blood sucking midge
Other morphological feature realizes, once encounter sibling species and morphological feature grasp it is very intractable not full Specimen identification；Moreover, right
The Morphological Identification of blood sucking midge needs to complete by making slide sample, complex for operation step, the period is long, difficulty is big, therefore, urgently
It need to seek a kind of new method, to make up the defect of conventional sorting methods.
Molecular Identification technology is to illustrate the difference between species so that hereditary material DNA sequence analysis is foundation, from molecular water
Quickly and accurately differentiate species on flat.It is the product that molecular biology, computer science are combined with traditional taxonomy, is made
For a kind of brand-new taxology technology, the defect of traditional form identification is greatly compensated for, more and more biologists have been caused
Attention.In recent years, with the foundation of the determined dna sequence method on the bases PCR and being widely used, rDNA
（rDNA）And mitochondrial DNA（mtDNA）Equal genes are gradually taken seriously, and are answered extensively in the Molecular Identification of blood sucking midge
With.The technology is proved each other with traditional sorting technique, can not only solve the problems such as sample is incomplete in identification, but also can be real
The Rapid identification of existing blood sucking midge.Thailand's Storehouse midge special gene sequence provided by the invention is advantageously implemented the quick mirror of Thailand's Storehouse midge
It is fixed, shorten qualification time.
Invention content
The purpose of the present invention is to provide a kind of special gene sequences and its method for identifying molecules of Thailand's Storehouse midge.By dividing
Sub- biological method obtains Thailand's Storehouse midge（Culicoides inthanonensis）Specific gene -16S rDNA gene orders,
By the comparison of gene order similitude, Thailand's Storehouse midge can be accurately and rapidly identified.
To achieve the above object, the present invention is achieved through the following technical solutions：
Using small insects minim DNA method for preparing template, by improved PCR reaction conditions, by the primer synthesized
The 16S rDNA genes of Thailand's Storehouse midge are expanded, PCR product sequence is sent to be measured by professional biotech firm.Sequencing result passes through manual school
To, sequence assembly, and Blast similarity searchings are carried out in NCBI, it is ensured that gained sequence is target sequence.It is of the present invention
Thailand's Storehouse midge specific gene, be 16S rDNA genes, the 16S rDNA gene orders such as SEQ ID No.1 institutes of the Storehouse midge
Show（Without primer）：
The PCR primer of Thailand's Storehouse midge 16S rDNA genes of the present invention, it is characterised in that PCR primer sequence is：
5 ' GCCTGTTTATCAAAAACAT 3 ' of forward primer
5 ' CCGGTCTGAACTCAGATCA 3 ' of reverse primer.
The method for identifying molecules of Thailand's Storehouse midge of the present invention, including：
1）The separation and Extraction DNA from insect tissue to be measured；
2）Using the DNA as template, the primer that uses for：5 ' GCCTGTTTATCAAAAACAT 3 ' of forward primer, reversely draws
5 ' CCGGTCTGAACTCAGATCA 3 ' of object goes out the 16S rDNA genes of the insect by PCR amplification；
3）Then appropriate step 2 is taken）The PCR amplification 16S rDNA gene agarose electrophoresis that goes out point
From, observed under ultraviolet lamp after ethidium bromide staining, according to the size of amplified production judge result.If can correspondingly specificity expand
Increase the band for that size is about 511 bp, biotech firm is sent to be sequenced；
4）According to sequencing result, if the similitude of target gene sequence and SEQ ID No.1 are 98% or more, you can judge
The test serum is Thailand's Storehouse midge.
The Identification of Species of Thailand's Storehouse midge is realized by Morphological Identification, and is checked through authoritative expert, to ensure to finish
The reliability of fruit.
The primer is synthesized according to document, 5 ' GCCTGTTTATCAAAAACAT 3 ' of forward primer, reverse primer 5 '
CCGGTCTGAACTCAGATCA 3’。
The present invention can use agarose gel electrophoresis detection method to detect amplification.
Advantages of the present invention is：
1）The present invention is combined using traditional Morphological Identification with molecular classification method, is identified the Storehouse midge,
It can ensure that the accuracy and reliability of species identification.
2）Compared with traditional Morphological Identification method, Thailand's Storehouse midge method for identifying molecules that the present invention establishes can effectively contract
The qualification time of short Thailand's Storehouse midge.
Description of the drawings
Fig. 1 is Thailand's Storehouse midge 16S rDNA gene PCR amplification electroresis appraisal figures.Number is described as follows：Swimming lane 1 is the moon
Property control, swimming lane 2-4 be Thailand's Storehouse midge female adult 16S rDNA genes, detection size be about 511 bp band.M is DL
2000 DNA marker.
Fig. 2 is unknown Storehouse midge female adult 16S rDNA gene PCR amplification electroresis appraisal figures.Number is described as follows：Swimming lane 1
For negative control, swimming lane 2-3 is the 16S rDNA genes of unknown Storehouse midge female adult, the band that detection size is about 511 bp.M is DL
2000 DNA marker.
Specific implementation mode
1 Thailand's Storehouse midge of embodiment（Culicoides inthanonensis）The acquisition of 16S rDNA gene orders
1, the acquisition of Thailand's Storehouse midge sample
Thailand's Storehouse midge is the sample that field acquisition obtains, and is directly stored in 95% alcohol after chilled execution.Sample is made
It is identified through authoritative expert after slide, it is ensured that the accuracy of qualification result.Before preparation of specimen, picking Thailand Storehouse midge chest end and abdomen
The former sections in portion are placed in 95% alcohol, are extracted for DNA.
2, prepared by DNA profiling
Thailand Storehouse midge DNA is extracted using small insects trace DNA templet preparation method（Wen Lizhang edits entomology research
Method and the Beijing technology introduction [M]：Science Press, 2010. pp278-286）, the DNA sample of extraction is stored in -20 DEG C
It is spare.
3, primer synthesizes
The present embodiment the primer is as follows：
5 ' GCCTGTTTATCAAAAACAT 3 ' of forward primer
5 ' CCGGTCTGAACTCAGATCA 3 ' of reverse primer.
4, PCR amplification
The PCR reaction systems of the present embodiment are as shown in table 1.
1 Thailand's Storehouse midge 16S rDNA gene PCR reaction systems of table（50uL systems）
| Ingredient | Volume（uL） |
| Template | 2 |
| Primer 1 | 1 |
| Primer 2 | 1 |
| 10 × buffer solution | 5 |
| dNTPs | 1 |
| Taq archaeal dna polymerases | 2.5 |
| ddH2O | 37.5 |
The response procedures of the present embodiment are as shown in table 2.
2 Thailand's Storehouse midge 16S rDNA gene PCR response procedures of table
| Step | Reaction temperature（℃） | Time | Recurring number |
| Pre-degeneration | 94 | 3 min | — |
| Denaturation | 94 | 45 s | 35 |
| Annealing | 48 | 45 s | 35 |
| Extend | 72 | 60 s | 35 |
| Extend | 72 | 7 min | — |
PCR product is saved backup in 4 DEG C.
5, pcr amplification product detects
5 μ L pcr amplification products are taken, with 1% agarose gel electrophoresis（130V voltages, 35 min of electrophoresis）.Ethidium bromide staining,
Gel imaging system detects, and electrophoresis pattern is referring to attached drawing 1.
6, gene sequencing
The selection preferable pcr amplification product of 3-5 effect is sent to be sequenced after purification by biotech firm（Be sequenced the primer with
PCR the primers are identical）, it is 16S rDNA gene orders-SEQ of Thailand's Storehouse midge after the calibrated splicing of gained gene order
ID No.1。
The identification of 2 unknown Storehouse midge of embodiment
1, the acquisition and preservation of sample
Using net method is waved or lamp lures method to be acquired, directly it is stored in 95% alcohol after the chilled execution of sample of harvest.
2, prepared by DNA profiling
Under anatomical lens or stereomicroscope, chooses a female Storehouse midge at random from collected midge class sample, use
Small insects trace DNA templet preparation method extracts Storehouse midge DNA（Wen Lizhang edits entomology research method and technology introduction
The Beijing [M]：Science Press, 2010. pp278-286）, the DNA sample of extraction saves backup in -20 DEG C, every part of sample one
A number.
3, the acquisition of objective gene sequence
3.1 primers synthesize
Using the unknown Storehouse midge DNA of acquisition as template, synthetic primer, the primer sequence is as follows：
5 ' GCCTGTTTATCAAAAACAT 3 ' of forward primer
5 ' CCGGTCTGAACTCAGATCA 3 ' of reverse primer.
3.2 PCR amplification
The PCR reaction systems of the present embodiment are as shown in table 3.
3 unknown Storehouse midge 16S rDNA gene PCR reaction systems of table（50uL systems）
| Ingredient | Volume（uL） |
| Template | 2 |
| Primer 1 | 1 |
| Primer 2 | 1 |
| 10 × buffer solution | 5 |
| dNTPs | 1 |
| Taq archaeal dna polymerases | 2.5 |
| ddH2O | 37.5 |
The response procedures of the present embodiment are as shown in table 4.
4 unknown Storehouse midge 16S rDNA gene PCR response procedures of table
| Step | Reaction temperature（℃） | Time | Recurring number |
| Pre-degeneration | 94 | 3 min | — |
| Denaturation | 94 | 45 s | 35 |
| Annealing | 48 | 45 s | 35 |
| Extend | 72 | 60 s | 35 |
| Extend | 72 | 7 min | — |
PCR product is saved backup in 4 DEG C.
3.3 pcr amplification products detect and gene sequencing
5 μ L pcr amplification products are taken, with 1% agarose gel electrophoresis（130V voltages, 35 min of electrophoresis）.Ethidium bromide contaminates
Color.Gel imaging system detects, and electrophoresis pattern is referring to attached drawing 2.Find out that the unknown Storehouse midge of embodiment 2 also can specificity by attached drawing 2
Amplify the product that size is about 511 bp, the product for being about 511 bp by size send biotech firm to be sequenced, as a result display with
The similitude of gene order SEQ ID NO. 1 is 99.9%, thus can determine that the unknown Storehouse midge is Thailand's Storehouse midge.
Claims (2)
1. a kind of Thailand's Storehouse midge specific gene, which is characterized in that the gene is 16S rDNA genes, is gene as described below
Sequence SEQ ID No.1：
。
2. a kind of method for identifying molecules of Thailand's Storehouse midge, including：
1）The separation and Extraction DNA from insect tissue to be measured；
2）Using the DNA as template, primer that 16S rDNA genes are used for：5 ' GCCTGTTTATCAAAAACAT of forward primer
3 ' ', 5 ' CCGGTCTGAACTCAGATCA 3 ' of reverse primer goes out Thailand Storehouse midge 16S rDNA by PCR amplification
Gene；
3）Then appropriate step 2 is taken）The 16S rDNA genes that go out of PCR amplification detached with agarose electrophoresis, pass through
It is observed under ultraviolet lamp after ethidium bromide staining, according to the judgement of the size of amplified production as a result, if energy specific amplification goes out size
For the band of 511 bp, biotech firm is sent to be sequenced；
4）According to sequencing result, if accordingly 16S rDNA gene orders and the similitude of SEQ ID No.1 are 98% or more, i.e.,
It can determine whether that the test serum is Thailand's Storehouse midge.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610012014.4A CN105543364B (en) | 2016-01-08 | 2016-01-08 | A kind of specific gene and its method for identifying molecules of Thailand's Storehouse midge |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610012014.4A CN105543364B (en) | 2016-01-08 | 2016-01-08 | A kind of specific gene and its method for identifying molecules of Thailand's Storehouse midge |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105543364A CN105543364A (en) | 2016-05-04 |
| CN105543364B true CN105543364B (en) | 2018-08-28 |
Family
ID=55822921
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610012014.4A Expired - Fee Related CN105543364B (en) | 2016-01-08 | 2016-01-08 | A kind of specific gene and its method for identifying molecules of Thailand's Storehouse midge |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105543364B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108486121B (en) * | 2018-01-25 | 2020-02-11 | 遵义医学院 | Specific DNA sequence of Cuicoides spinuloides and molecular identification method thereof |
| CN107988232A (en) * | 2018-01-25 | 2018-05-04 | 遵义医学院 | The DNA bar code standard sequence and its method for identifying molecules of a kind of open country Storehouse midge |
| CN108034733B (en) * | 2018-01-29 | 2020-05-05 | 遵义医学院 | DNA bar code standard sequence of Culicoides kindred species and molecular identification method thereof |
| CN108486122A (en) * | 2018-01-29 | 2018-09-04 | 遵义医学院 | A kind of the DNA bar code standard sequence and its method for identifying molecules of South Mountain Storehouse midge |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102690829B (en) * | 2012-05-16 | 2013-09-25 | 福建国际旅行卫生保健中心 | A DNA bar code standard gene sequence for Taiwan Lasiohelea |
| KR101443716B1 (en) * | 2014-04-08 | 2014-09-26 | 주식회사 현일바이오 | Methods for Simultaneously Detecting Nontuberculous mycobacteria and Kits Using the Same |
-
2016
- 2016-01-08 CN CN201610012014.4A patent/CN105543364B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN105543364A (en) | 2016-05-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105543249B (en) | A kind of beauty Storehouse midge specific gene and its method for identifying molecules | |
| CN105483261B (en) | A kind of specific gene and its method for identifying molecules of indigo plant abdomen Storehouse midge | |
| CN105543364B (en) | A kind of specific gene and its method for identifying molecules of Thailand's Storehouse midge | |
| CN103805609B (en) | A kind of lasiohelea taiwana molecular assay method | |
| CN107557478A (en) | A kind of method and its application that amplification the Changjiang river fish chondriogen total order is combined based on degenerate primer | |
| CN107326077A (en) | A kind of molecular labeling for differentiating spotted maigre genetic sex and its application | |
| CN106086167B (en) | The primer sequence and method of a kind of Rapid identification Larimichthys crocea, little yellow croaker and Collichthys lucidus | |
| CN111471756B (en) | Specific DNA fragment SSM1 for sturgeon gender identification and application | |
| CN110607359A (en) | Patinopecten yessoensis female specific marker combination and application | |
| CN105420399B (en) | A kind of specific gene and its method for identifying molecules of the sub- naked midge of palm fibre | |
| CN113637765A (en) | Molecular marker for identifying genetic sex of micropterus salmoides and application | |
| CN105505942B (en) | A kind of specific gene and its method for identifying molecules of peacefulness pincers midge | |
| CN103952420A (en) | Dasyhelea specific gene sequence | |
| CN108384791A (en) | A kind of specific gene and its method for identifying molecules of Beijing Storehouse midge | |
| CN106119384B (en) | Aeromonas hydrophila nucleic acid analysis method and application thereof in forensic detection | |
| CN110724735B (en) | SNP locus and primer for rapidly identifying individual sex of fugu obscurus and method thereof | |
| CN104498593B (en) | Identify or auxiliary identify storage bean weevil primer to and test kit | |
| CN108486121B (en) | Specific DNA sequence of Cuicoides spinuloides and molecular identification method thereof | |
| CN104962660B (en) | A kind of mottle clam species real-time fluorescence PCR specific detection system and application | |
| CN109810982A (en) | The specific gene and its method for identifying molecules of short hairs Storehouse midge | |
| CN113981108B (en) | Molecular marker for genetic sex identification of megalobrama amblycephala and application of molecular marker | |
| CN108342398A (en) | A kind of specific gene and its method for identifying molecules of open country Storehouse midge | |
| CN108486122A (en) | A kind of the DNA bar code standard sequence and its method for identifying molecules of South Mountain Storehouse midge | |
| CN104673890B (en) | Cotton black aphid PCR quick determination methods based on specific SS COI primers | |
| CN113881786A (en) | Primers, primer kit and identification method for identifying Pseudobagrus ussuriensis, Pseudobagrus ussuriensis and cross-crossing individuals of Pseudobagrus ussuriensis and Pseudobagrus ussuriensis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180828 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |