CN107653329A - For identifying Specific PCR primers and method and its application in barking deer is identified of barking deer - Google Patents
For identifying Specific PCR primers and method and its application in barking deer is identified of barking deer Download PDFInfo
- Publication number
- CN107653329A CN107653329A CN201711019250.XA CN201711019250A CN107653329A CN 107653329 A CN107653329 A CN 107653329A CN 201711019250 A CN201711019250 A CN 201711019250A CN 107653329 A CN107653329 A CN 107653329A
- Authority
- CN
- China
- Prior art keywords
- pcr primers
- specific pcr
- deer
- barking
- barking deer
- 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
Abstract
The invention discloses a kind of Specific PCR primers for being used to identify barking deer (Elaphodus cephalophus), wherein, the Specific PCR primers include such as SEQ ID No：Primer 1 shown in 1, and such as SEQ ID No：Primer 2 shown in 2.The invention also discloses a kind of method that barking deer is identified using Specific PCR primers, this method includes：Extract the STb gene of testing sample and enter performing PCR amplification to STb gene；Product after amplification is done into agarose gel electrophoresis detection；Wherein, the Specific PCR primers are above-mentioned Specific PCR primers, if occurring the band of 1 length-specific in swimming lane, judge the testing sample for the sample from barking deer.The invention also discloses a kind of application of above-mentioned Specific PCR primers or authentication method in barking deer is identified.By designing above-mentioned primer, the purpose for identifying barking deer simple and easy to do and economical and practically is realized.
Description
Technical field
The present invention relates to field of biological detection, in particular it relates to a kind of Specific PCR primers for being used to identify barking deer and
Use the method for Specific PCR primers identification barking deer, and their applications in barking deer is identified.
Background technology
Barking deer (Elaphodus cephalophus) is a kind of middle-size and small-size phytophage deer for moving in hilly and mountainous land area
Section animal, it is subordinate to Mammalia (Mammalia), Artiodactyla (Artiodactyla), Cervidae (Cervidae) in classification.Barking deer
It is distributed mainly on the ground such as Chinese Zhejiang, Fujian, Anhui, Jiangxi, Guangdong, Hunan, Hubei, Sichuan, Yunnan.Document is documented in state
Outer Upper Myanmar is also distributed, but does not find existence individual so far.Therefore, barking deer is considered a kind of special product in me
The rare animal in deer family of state.Such as other animal in deer family, barking deer species are also deep to be influenceed by illegal hunt unlawfully with illegal trade, its
Population quantity is in continuous decrease state.Promulgate within 2000《State guarantee beneficial has Important Economic, scientific research
The terrestrial wildlife register of value》Protection species are classified as, are newly issued within 2015《Chinese biological diversity red name
Record-vertebrate》Barking deer is more classified as Chinese easily danger (VU) kind.Increase the protection and biological study to barking deer species
The common recognition of vast animal protection worker is turned into.
It is primary premise based on taxonomic accurate species identification in the protection of wild animal and management work.Thing
The result of kind identification is often that the administrative law-enforcement departments such as Forest Police, industry and commerce, frontier inspection carry out wildlife conservation and management
The basis of work.Traditional morphological classification method is always the common method of wild animal identification, but morphological classification method
Some intrinsic defects also bring difficulty, such as phenotypical plasticity and genetic variability, biological sex to species identification work
With the stage of development limitation etc..Appraisal for animal in deer family practice have shown that, due to stealing value of the shooter to its hide and meat
Seize, cause submitted sample fragmentation (sample type is hair, deformed limb, bone, hide and excrement etc., and sample size is smaller),
It can not be realized from morphology and species are accurately identified.
As PCR (Polymerase Chain Reaction, PCR) technology is in Forensic Identification
Constantly application, also obtained extensively in endangered wildlife is identified by the Protocols in Molecular Biology of representative of DNA sequence analysis
Application.
As PCR (Polymerase Chain Reaction, PCR) technology is in Forensic Identification
Constantly application, is also obtained by the Protocols in Molecular Biology of representative of DNA sequence analysis in endangered wildlife species identification
It is widely applied.
Therefore it provides the tedious steps such as a kind of sequencing without DNA, sequence alignment, while it is unrestricted to sample source, and
Only can fast and effeciently be realized by a PCR amplification is to the Specific PCR primers of the accurate measure of barking deer species
The problem of urgent need to resolve of the present invention.
The content of the invention
For above-mentioned prior art, it is an object of the invention to overcome in the prior art by morphological feature to barking deer
The limitation that species are measured, there is provided a kind of simple and efficient and economic Specific PCR primers for being used to identify barking deer and
The method for identifying barking deer, and their applications in barking deer is identified.
To achieve these goals, the present invention provides a kind of for identifying barking deer (Elaphodus cephalophus)
Specific PCR primers, the Specific PCR primers include such as SEQ ID No：Primer 1 shown in 1, and such as SEQ ID
No：Primer 2 shown in 2.The specific primer can enter performing PCR amplification to target sample, according to agarose gel electrophoresis
Separating resulting, which whether there is, produces purpose amplified production, to identify species to be checked.
The present invention also provides a kind of method using specific PCR identification barking deer (Elaphodus cephalophus),
Methods described includes：(1) STb gene of testing sample is extracted；(2) enter performing PCR to the STb gene using Specific PCR primers to expand
Increase；(3) product obtained in step (2) is taken to do agarose gel electrophoresis detection；Wherein, the Specific PCR primers are right
It is required that the Specific PCR primers described in 1；Wherein, in the agarose gel electrophoresis figure obtained by step (3), if containing step
(2) occur 1 band in the swimming lane of the product obtained, then the testing sample is judged for the sample from barking deer, if containing
1 band is occurred without in the swimming lane for the product that step (2) obtains, then judges that the testing sample is not from the sample of barking deer
Product.
In the present invention, the length of DNA fragmentation contained in 1 band of appearance is 240bp.
In the present invention, in order to reach more preferable PCR expanding effects, so as to get agarose gel electrophoresis figure it is apparent can
See, be easy to preferably determine experimental result, in PCR amplification procedures, on the basis of 30 μ L PCR amplification system, primer it is dense
Degree is preferably 0.25-0.5pmol/L.
PCR amplification system refers to including primer, Taq enzyme, dNTPMix, DNA profiling (STb gene), Mg2+And buffer solution etc. enters
All reagents and raw material that performing PCR amplification procedure is added.
In addition, in the present invention, in order to reach more preferable PCR expanding effects, so as to get agarose gel electrophoresis figure more
It is high-visible, it is easy to preferably determine experimental result, PCR amplifications are 45-55ng with the dosage of STb gene in step (2).
Wherein, the archaeal dna polymerase and buffer solution for PCR amplifications can be this area routine archaeal dna polymerase and buffering
Liquid, its dosage are specifically referred to the detailed description in product description.In this not go into detail by the present invention.
In the present invention, in order to reach more preferable PCR expanding effects, so as to get agarose gel electrophoresis figure it is apparent can
See, be easy to preferably determine experimental result, it is preferable that PCR amplification procedures include 28-33 circulation, single loop include be denatured,
The step of annealing and extension.
Annealing process in the circular response can be the conventional use of annealing way in this area and annealing parameter,
In the present invention, in order to obtain more preferable expanding effect, annealing temperature is 55-65 DEG C, annealing time 25-40s.
Degenerative process in the circular response can be the conventional use of denaturation way in this area and denaturation parameter,
In the present invention, in order to obtain more preferable expanding effect, denaturation temperature is 93-95 DEG C.
Extension mode and extension parameter in the circular response, in the present invention, in order to obtain more preferable expanding effect,
Elongating temperature is 70-73 DEG C.
In addition, in order to ensure to obtain more preferable expanding effect, the amplification procedure also include before circulating first 93-95 DEG C it is pre-
5min, 58 DEG C of annealing temperature are denatured, last time circulation re-extends 10min after 71-73 DEG C.
Present invention also offers one kind according to above-mentioned Specific PCR primers or above-mentioned authentication method in identification barking deer
Application in (Elaphodus cephalophus).
The present invention, using the method for specific PCR amplification, passes through one by designing Specific PCR primers to testing sample
Secondary conventional PCR amplifications can be shown by the band in agarose gel electrophoresis figure determines whether the species are barking deer,
And DNA profiling used can extract from the fur of animal, musculature, internal organ, the difficulty of sampling, and the party are greatly reduced
Method can quickly and easily identify barking deer, so as to reach simple and easy to do and economical and practical the purpose for identifying barking deer.Its is whole
Individual detection process takes altogether is no more than 3 hours, and operating process is simple, is with a wide range of applications.The present invention relates to
Primer and authentication method developing into the identification that after kit, can be widely applied to barking deer species and population detection research
In, there is larger promotional value.
Specific embodiment part below is described in further detail advantages of the present invention.
Brief description of the drawings
Accompanying drawing is for providing a further understanding of the present invention, and a part for constitution instruction, with following tool
Body embodiment is used to explain the present invention together, but is not construed as limiting the invention.In the accompanying drawings：
Fig. 1 is the agarose gel electrophoresis figure of embodiment 1-4 and comparative example 1-11 amplified production.
Embodiment
The embodiment of the present invention is described in detail below.It is it should be appreciated that described herein specific
Embodiment is merely to illustrate and explain the present invention, and is not intended to limit the invention.
The present invention will be described in detail by way of examples below.
It is necessary to clarify that the primer is synthesized by Shanghai Sheng Gong biotechnologies Co., Ltd and primer is using dense
It can be DNA lysates commonly used in the art to spend for 10pmol/L, the lysate, for example, in the present invention, every liter of institute
State Tris-HCl (pH8.0), 25mmol ethylenediamine tetra-acetic acid (pH8.0), the 100mmol chlorine in lysate containing 50mmol
Change the lauryl sodium sulfate of sodium and 1% parts by weight, the dNTPMix used in the present invention is limited for the full formula gold biotechnology in Beijing
The commercially available product of company, Proteinase K are the commercially available product of Merck companies of the U.S., and the DNA purification kits are that Beijing Tiangeng is biochemical public
The commercially available product of department, remaining used chemical reagent are conventional commercial AR.PCR is reacted in 2720 type PCR instruments
Carried out in (ABI, the U.S.)；Agarose gel electrophoresis testing result by HL-2000 types gel imaging system (UVP, the U.S.) analyze,
Take pictures.
Embodiment 1
Using the method for Specific PCR primers identification barking deer, including：
(1) STb gene of testing sample is extracted：
The testing sample that numbering is EC1 (being shown in Table 1) in 0.5g tables 1 is placed in centrifuge tube and test sample will be treated in centrifuge tube
Product are shredded with sterilization scissors, and 500 μ L lysate, 30 μ L 10% lauryl sodium sulfate are then sequentially added into centrifuge tube
(SDS) and 3 μ L 20mg/ml Proteinase K (PK), it is the final concentration of 100 μ g/ul of 0.5%, PK to make SDS concentration, fully mix
12h is digested to liquid in pipe clarification after 56 DEG C of water-baths.The μ L of balance phenol 500 are added into above-mentioned postdigestive mixture, and gently
It is placed in after micro oscillation 5min on 11000r/min centrifuge and centrifuges 10min, takes the supernatant after centrifugation.Repeat above-mentioned centrifugation step
Suddenly twice.Add two mixed (chloroforms：Isoamyl alcohol=24：1) 500 μ l, slight oscillatory 5min, 11000rpm centrifugation 10min, afterwards
Supernatant is taken to be transferred in new EP pipes.Two steps more than repeating.Frost nothing is added in the supernatant finally given after to repeated centrifugation
The μ L of water-ethanol 1000 and at -20 DEG C place 1h after be placed on 12000r/min centrifuge centrifuge 13min after, abandon supernatant,
And the 70% μ L of ethanol 800 are added into obtained precipitation, and 13000r/min centrifuge is placed in after slight oscillatory 0.5min
Upper frost centrifugation 13min, abandons supernatant.Repeat above-mentioned centrifugation step twice.Obtained precipitation is placed on aseptic operating platform certainly
So dry and add the μ L of TE solution 300 after 2.5h thereto, slight oscillatory simultaneously flicks centrifuge tube after placing 3h at 4 DEG C with finger
Afterwards, STb gene is obtained；
(2) performing PCR amplification is entered to the STb gene using Specific PCR primers：
10 μ L 10 × PCR buffer, 10pmol/L each 1 μ L of one couple of PCR primers are added into the sample cell of PCR instrument,
2 μ L 2mmol/L dNTPMix, 2 μ L 25mmol/L Mg2+, 1U Taq enzyme and 50ng STb gene, and mended with distilled water
Together to 30 μ L.PCR reaction conditions are：95 DEG C of pre-degeneration 5min；Then 30 circulations are carried out, the circulation includes：95 DEG C of denaturation
40s, 30s and 72 DEG C of extension 35s of 60 DEG C of annealing；Finally remake 72 DEG C of extension 10min.Enter performing PCR expansion under the above-described reaction conditions
Increase；
(3) product obtained in step (2) is taken to do agarose gel electrophoresis detection：
Amplified production electrophoresis detection on 1% Ago-Gel through EB dyeing, keep 5V/cm voltage 30min, Yu Zi
Observed on outer gel imaging system, shooting preserves.Electrophoresis result is as shown in figure 1, corresponding swimming lane numbering is 7.
Embodiment 2
Method according to embodiment 1 is operated, unlike, 10pmol/L one couple of PCR primers each 0.75 μ L are described
The dosage of STb gene is 45ng, and PCR amplification procedures include 28 circulations, and denaturation temperature is 93 DEG C in single loop, and annealing temperature is
55 DEG C, annealing time 25s, elongating temperature is 70 DEG C.By the DNA after amplification 1% through EB dyeing Ago-Gel on electricity
Swimming detection, keeps 5V/cm voltage 30min, is observed on ultraviolet gel imaging system, and electrophoresis result is shown in 240bp appearance
One band.
Embodiment 3
Method according to embodiment 1 is operated, unlike, 10pmol/L one couple of PCR primers each 1.5 μ L are described
The dosage of STb gene is 55ng, and PCR amplification procedures include 33 circulations, and denaturation temperature is 95 DEG C in single loop, and annealing temperature is
65 DEG C, annealing time 40s, elongating temperature is 73 DEG C.By the DNA after amplification 1% through EB dyeing Ago-Gel on electricity
Swimming detection, keeps 5V/cm voltage 30min, is observed on ultraviolet gel imaging system, and electrophoresis result is shown in 240bp appearance
One band.
Embodiment 4
Method according to embodiment 1 is operated, unlike, testing sample numbering is EC2 (in table 1), electrophoresis result
As shown in figure 1, corresponding swimming lane numbering is 6.
Comparative example 1
Method according to embodiment 1 is operated, unlike, testing sample numbering is OG (in table 1), and electrophoresis result is such as
Shown in Fig. 1, corresponding swimming lane numbering is 1.
Comparative example 2
Method according to embodiment 1 is operated, unlike, testing sample numbering is BK (in table 1), and electrophoresis result is such as
Shown in Fig. 1, corresponding swimming lane numbering is 3.
Comparative example 3
Method according to embodiment 1 is operated, unlike, testing sample numbering is SE (in table 1), and electrophoresis result is such as
Shown in Fig. 1, corresponding swimming lane numbering is 2.
Comparative example 4
Method according to embodiment 1 is operated, unlike, testing sample numbering is MT (in table 1), and electrophoresis result is such as
Shown in Fig. 1, corresponding swimming lane numbering is 5.
Comparative example 5
Method according to embodiment 1 is operated, unlike, testing sample numbering is SS (in table 1), and electrophoresis result is such as
Shown in Fig. 1, corresponding swimming lane numbering is 10.
Comparative example 6
Method according to embodiment 1 is operated, unlike, testing sample numbering is CS (in table 1), and electrophoresis result is such as
Shown in Fig. 1, corresponding swimming lane numbering is 11.
Comparative example 7
Method according to embodiment 1 is operated, unlike, testing sample numbering is MB (in table 1), and electrophoresis result is such as
Shown in Fig. 1, corresponding swimming lane numbering is 4.
Comparative example 8
Method according to embodiment 1 is operated, unlike, testing sample numbering is HI (in table 1), and electrophoresis result is such as
Shown in Fig. 1, corresponding swimming lane numbering is 8.
Comparative example 9
Method according to embodiment 1 is operated, unlike, testing sample numbering is CN (in table 1), and electrophoresis result is such as
Shown in Fig. 1, corresponding swimming lane numbering is 9.
Comparative example 10
Method according to embodiment 1 is operated, unlike, testing sample numbering is MC (in table 1), and electrophoresis result is such as
Shown in Fig. 1, corresponding swimming lane numbering is 12.
Comparative example 11
Method according to embodiment 1 is operated, unlike, testing sample numbering is MM (in table 1), and electrophoresis result is such as
Shown in Fig. 1, corresponding swimming lane numbering is 13.
Reference examples 1
To avoid external source from polluting, the operation in the STb gene and PCR amplification procedures of all extraction testing samples all sets sterile double
The negative control that water is template is steamed, electrophoresis result is as shown in figure 1, corresponding swimming lane numbering is C.
Table 1
As shown in table 1 and Fig. 1, swimming lane is respectively in Fig. 1, M：D2000 molecular labelings (Tiangeng is biochemical)；1：OG；2：
SE；3：BK；4：MB；5：MT；6：EC2；7：EC1；8：HI；9：CN；10：SS；11：CS；12：
MC；13：MM；C roads：Blank control.The present invention is can be seen that by the electrophoretogram shown in the sample message and Fig. 1 of table 1 to pass through
1 group-specific PCR primer is designed, agarose gel electrophoresis only can be passed through by once conventional PCR amplifications to testing sample
Band in figure, which is shown, determines whether the species are barking deer, and electrophoresis result complies fully with sample message, it was confirmed that should
The reliability of authentication method.
Detect example 1
The Ago-Gel block containing target DNA fragment in band will be obtained to be cut with clean scalpel and pure in DNA
Shanghai Sheng Gong biotechnologies Co., Ltd is delivered to after purification in change kit to be sequenced.Shanghai life work biology work will be delivered to
The sequence for the DNA fragmentation that journey Technology Co., Ltd. is sequenced and the sequence of known barking deer species in GenBank databases
After being compared, obtained comparison result shows the sequence of the DNA fragmentation of the sequencing and the sequence of known barking deer species
Homology is more than 96%, so the species that also further demonstrate the testing sample are barking deer.
Moreover, to ensure the reliability of experimental result, we enter the amplified production to barking deer sample EC1 and EC2
Sequencing is gone.Using GenBank databases Blast software searches compare after find, the sequence of EC1 and EC2 samples with
The sequence similarity of existing barking deer species homologous fragment sequence has reached 100% in GenBank, and its GenBank is logged in
Number be respectively KU324462, KU324463 and KU324464.This shows the PCR specific primers of the present invention, can be effectively right
Barking deer species realize specific amplification.
In the present invention, DNA profiling used can extract from the musculature of animal, greatly reduce the difficulty of sampling
Degree, and this method can quickly and easily identify barking deer, and barking deer is identified so as to reach simple and easy to do and economical and practical
Effect.
The preferred embodiment of the present invention described in detail above, still, the present invention are not limited in above-mentioned embodiment
Detail, in the range of the technology design of the present invention, a variety of simple variants can be carried out to technical scheme, this
A little simple variants belong to protection scope of the present invention.
It is further to note that each particular technique feature described in above-mentioned embodiment, in not lance
In the case of shield, can be combined by any suitable means, in order to avoid unnecessary repetition, the present invention to it is various can
The combination of energy no longer separately illustrates.
In addition, various embodiments of the present invention can be combined randomly, as long as it is without prejudice to originally
The thought of invention, it should equally be considered as content disclosed in this invention.
SEQUENCE LISTING
<110>Anhui Normal University
<120>For identifying Specific PCR primers and method and its application in barking deer is identified of barking deer
<130> 05631
<160> 2
<170> PatentIn version 3.3
<210> 1
<211> 20
<212> DNA
<400> 1
tactctacca aatctctccg 20
<210> 2
<211> 27
<212> DNA
<400> 2
aaaagggtaa tgagggctaa gacggtc 27
Claims (10)
1. one kind is used for the Specific PCR primers for identifying barking deer (Elaphodus cephalophus), it is characterised in that institute
Stating Specific PCR primers includes such as SEQ ID No：Primer 1 shown in 1, and such as SEQ ID No：Primer 2 shown in 2.
A kind of 2. method that barking deer is identified using Specific PCR primers, it is characterised in that methods described includes：
(1) STb gene of testing sample is extracted；
(2) performing PCR amplification is entered to the STb gene using Specific PCR primers；
(3) product obtained in step (2) is taken to do agarose gel electrophoresis detection；
Wherein, the Specific PCR primers are the Specific PCR primers described in claim 1；
Wherein, in the agarose gel electrophoresis figure obtained by step (3), if going out in the swimming lane of the product obtained containing step (2)
Existing 1 band, then judge the testing sample for the sample from barking deer, if the swimming lane of the product obtained containing step (2)
In occur without 1 band, then judge that the testing sample is not from the sample of barking deer.
3. according to the method for claim 2, wherein, the length of contained DNA fragmentation is in 1 band of appearance
240bp。
4. according to the method in claim 2 or 3, wherein, in PCR amplification procedures, using 30 μ L PCR amplification system as base
Standard, the concentration of the primer is 0.25-0.5pmol/L.
5. according to the method for claim 4, wherein, PCR amplifications are 45-55ng with the dosage of STb gene in step (2).
6. according to the method in claim 2 or 3, wherein, PCR amplification procedures include 28-33 circulation, single loop includes
The step of denaturation, annealing and extension.
7. according to the method for claim 6, wherein, annealing temperature is 55-65 DEG C, annealing time 25-40s.
8. according to the method for claim 6, wherein, denaturation temperature is 93-95 DEG C.
9. according to the method for claim 6, wherein, elongating temperature is 70-73 DEG C.
10. Specific PCR primers according to claim 1 or the method any one of claim 2-9 are being identified
Application in barking deer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711019250.XA CN107653329B (en) | 2017-10-27 | 2017-10-27 | Specific PCR primer and method for identifying hairy deer and application of specific PCR primer and method in identifying hairy deer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711019250.XA CN107653329B (en) | 2017-10-27 | 2017-10-27 | Specific PCR primer and method for identifying hairy deer and application of specific PCR primer and method in identifying hairy deer |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107653329A true CN107653329A (en) | 2018-02-02 |
| CN107653329B CN107653329B (en) | 2021-05-14 |
Family
ID=61095163
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711019250.XA Active CN107653329B (en) | 2017-10-27 | 2017-10-27 | Specific PCR primer and method for identifying hairy deer and application of specific PCR primer and method in identifying hairy deer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107653329B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050233334A1 (en) * | 2004-04-16 | 2005-10-20 | Toyoko Kusama | Oligonucleotide sequences that identify species of animal |
| CN1932041A (en) * | 2005-07-06 | 2007-03-21 | 韩国韩医学研究院 | The primers specific to cervus elaphus, c. nippon, c. canadensis and rangifer tarandus gene and the method to identify cervi parvum cornu species |
| CN104946788A (en) * | 2015-07-25 | 2015-09-30 | 舟山市食品药品检验检测研究院 | PCR primer and kit for identifying eight kinds of animal-derived components |
| CN105441537A (en) * | 2015-11-05 | 2016-03-30 | 陕西省动物研究所 | Method for molecular identification of Qinling Mountain illegal trade animal species and primer thereof |
| CN106011276A (en) * | 2016-07-18 | 2016-10-12 | 安徽师范大学 | Species-specificity PCR (polymerase chain reaction) identifying primer and method for identifying glandirana tientaiensis and application |
| CN106434921A (en) * | 2016-09-29 | 2017-02-22 | 中国科学院城市环境研究所 | Microbial source tracking molecular marker and high-throughput detection method thereof for detecting multiple fecal pollution sources |
-
2017
- 2017-10-27 CN CN201711019250.XA patent/CN107653329B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050233334A1 (en) * | 2004-04-16 | 2005-10-20 | Toyoko Kusama | Oligonucleotide sequences that identify species of animal |
| CN1932041A (en) * | 2005-07-06 | 2007-03-21 | 韩国韩医学研究院 | The primers specific to cervus elaphus, c. nippon, c. canadensis and rangifer tarandus gene and the method to identify cervi parvum cornu species |
| CN104946788A (en) * | 2015-07-25 | 2015-09-30 | 舟山市食品药品检验检测研究院 | PCR primer and kit for identifying eight kinds of animal-derived components |
| CN105441537A (en) * | 2015-11-05 | 2016-03-30 | 陕西省动物研究所 | Method for molecular identification of Qinling Mountain illegal trade animal species and primer thereof |
| CN106011276A (en) * | 2016-07-18 | 2016-10-12 | 安徽师范大学 | Species-specificity PCR (polymerase chain reaction) identifying primer and method for identifying glandirana tientaiensis and application |
| CN106434921A (en) * | 2016-09-29 | 2017-02-22 | 中国科学院城市环境研究所 | Microbial source tracking molecular marker and high-throughput detection method thereof for detecting multiple fecal pollution sources |
Non-Patent Citations (3)
| Title |
|---|
| CAO,X.ET AL: "Elaphodus cephalophus mitochondrion, complete genome", 《GENBANK》 * |
| HONG PANG ET AL: "Identification of complete mitochondrial genome of the tufted deer", 《DNA SEQUENCE》 * |
| 徐佳萍 等: "线粒体DNA在鹿科动物物种鉴定中的应用", 《草食家畜》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107653329B (en) | 2021-05-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| ES2555389T3 (en) | Analysis of gene expression in individual cells | |
| EP2115123A2 (en) | Selective lysis of sperm cells | |
| KR20240046529A (en) | Method for species identification of eukaryotes based on whole genome analysis and uses thereof | |
| CN105441421A (en) | Cell lysis method | |
| CN108913768B (en) | Multiplex liquid phase gene chip primer, kit and analysis method for simultaneously detecting seven glucosamine drug resistance genes | |
| CN105441547B (en) | For identifying multiple PCR primer and method and the application of Anji Chinese salamander | |
| Koh et al. | Extraction of high molecular weight DNA suitable for next-generation sequencing from the fiber crop abaca | |
| Delalande et al. | Multigenic families and proteomics: extended protein characterization as a tool for paralog gene identification | |
| El-Matbouli et al. | Detection of Cyprinid herpesvirus-3 (CyHV-3) DNA in infected fish tissues by nested polymerase chain reaction | |
| CN107653329A (en) | For identifying Specific PCR primers and method and its application in barking deer is identified of barking deer | |
| CN111690759A (en) | Specific primer, kit and method for detecting RPA of citrus canker pathogen | |
| CN116622909A (en) | Isothermal amplification detection reagent and detection method for feline herpesvirus I type | |
| CN111763748B (en) | Universal single primer, kit and identification method for identifying tridacna, tridacna without scales and trioyster | |
| CN103409515A (en) | Kit used for species identification of channel catfish and application method of kit | |
| CN112795662A (en) | Identification method of barbel grahami and application thereof | |
| CN103243089B (en) | Method for extracting deoxyribonucleic acid (DNA) from tanned animal specimen hide | |
| CN106834489A (en) | For identifying the Specific PCR primers and method of black muntjac and its application in black muntjac is identified | |
| CN109652520B (en) | Composition for detecting conversion efficiency of bisulfite-treated DNA in DNA methylation assays | |
| CN107287315B (en) | Detection kit for Shigella, detection method and application | |
| CN104372074B (en) | The reagent of a kind of energy specific detection Mia antigen gene and detection method | |
| CN104059979B (en) | For the identification of the multiple PCR primer of leopard and method and their application in qualification leopard | |
| CN107475433A (en) | One kind mixing animal species authentication method and primer | |
| CN106755536A (en) | Specific PCR primers and the method using Specific PCR primers identification serow for identifying serow | |
| CN111455069B (en) | Method for rapidly identifying DNA of ivory and ivory product | |
| CN111763747B (en) | Universal single primer, kit and identification method suitable for identifying common giant clam species in south sea area |
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 |