CN109136338A - A kind of primer, probe and kit and method detecting genetically engineered soybean MON89788 - Google Patents
A kind of primer, probe and kit and method detecting genetically engineered soybean MON89788 Download PDFInfo
- Publication number
- CN109136338A CN109136338A CN201811222863.8A CN201811222863A CN109136338A CN 109136338 A CN109136338 A CN 109136338A CN 201811222863 A CN201811222863 A CN 201811222863A CN 109136338 A CN109136338 A CN 109136338A
- Authority
- CN
- China
- Prior art keywords
- primer
- probe
- genetically engineered
- engineered soybean
- detection
- 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
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
- C12Q1/6844—Nucleic acid amplification reactions
-
- 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
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Botany (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of primer, probe and kits and method for detecting genetically engineered soybean MON89788.Primer sets are made of 2 specific primers, and nucleotide sequence is as shown in No:1~2 SEQ ID, and probe sequence is as shown in SEQ ID No.3.Detection kit includes detection primer and probe solution, Buffer A, Buffer B and positive control.Detection method is to use specific primer and probe under the action of recombinase, single-stranded DNA binding protein (SSB), strand displacement archaeal dna polymerase and exonuclease, sample DNA templates are expanded at 37~42 DEG C, by the method to fluorescence probe signal collection, whether judgement sample contains genetically engineered soybean MON89788 ingredient.The present invention does not need specific apparatus, have the characteristics that rapidly and efficiently, easy to operate, high sensitivity, high specificity, be suitble to on-site test.
Description
Technical field
The invention belongs to technical field of molecular biology, are related to the detection method of genetically modified plants and products thereof, specifically relate to
And it is a kind of quick using recombinase polymeric enzymatic amplification technology (Recombinase Polymerase Amplification, RPA)
Detect the primer and probe combinations, kit and detection method of genetically engineered soybean MON89788.
Background technique
Global genetically modified crops cultivated area is up to 1.851 hundred million hectares within 2016, and than 1996 commercialization initial stages increased
More than 100 times.The genetically modified crops of commercial growth mainly have soybean, corn, cotton, rape at present, wherein grown worldwide area
Maximum is genetically engineered soybean, reaches 91,400,000 hectares within 2016, accounts for the 50% of the global genetically modified crops gross area.Transgenosis is big
Beans MON89788 is a kind of Herbicide-Resistant Transgenic of Monsanto Chemicals' research and development, is used by Chinese government's approval of import
It processes raw material.The detection method research for carrying out fast accurate for genetically engineered soybean MON89788, is GMO bio-safety
The technical support and guarantee that management relevant laws and regulations are smoothly implemented.
Detection of GMOs technology is broadly divided into two major classes: one kind is using exogenous DNA as test object, such as PCR, gene core
Piece etc., it is another kind of using the protein of exogenous gene expression as test object, such as ELISA, immunity test strip.In the above method
In, round pcr is most widely used at present, but the transgenic detection method of based on PCR needs PCR instrument, gel imaging system etc.
Special instrument equipment, and expand longer (about 3~4h) with the product detection time, it is difficult to reach field quick detection purpose, therefore,
A kind of detection GMOs new technology that is more convenient, accurate and being suitable for execute-in-place is needed in actual operation.
RPA technology is a kind of novel gene amplification technology, depends on three kinds of enzymes: can combine single-chain nucleic acid (few core
Thuja acid primer) recombinase, single-stranded DNA binding protein (SSB) and strand displacement archaeal dna polymerase.The mixture of these three enzymes is normal
Also active under temperature, optimal reaction temperature is at 37 DEG C or so.The Protein-DNA mixtures that recombinase is formed in conjunction with primer, can be
Homologous sequence is found in double-stranded DNA.Once primer located homologous sequence, Exchange reaction of chain will occur and in strand displacement DNA
It polymerize starting DNA synthesis under enzyme effect, exponential amplification is carried out to the target area in template, the DNA chain being replaced and SSB are tied
It closes, prevents from further replacing.In this system, a compound event is originated by two opposite primers.Whole process carries out
Must be very fast, can generally obtain within ten minutes can detect horizontal amplified production.The basic characteristics of the technology are: 1. constant temperature
Amplification: entire amplified reaction is carried out at (37~42 DEG C) of constant temperature, does not need special instruments and equipment；2. rapidly and efficiently: entire
Amplification and product detection can be completed in 30 minutes；3. high specific: recombinase and primer form complex, special searching target
Sequence is marked, along with the specific recognition and signal collection of probe, specific amplification is high；4. highly sensitive: detectable limit can be down to
10 copies or lower；5. identification is easy: by the presence or absence of amplification fluorescent curve, easily being detected to amplified production.
RPA method have the characteristics that rapidly and efficiently, easy to operate, high specificity, high sensitivity, do not need specific apparatus,
It is suitble to field quick detection, has broad application prospects in transgenic plant detection field.There has been no utilize RPA method at present
Detect the detection method and kit of genetically engineered soybean MON89788.
Summary of the invention
The purpose of the present invention is intended to provide the primer and probe of RPA detection genetically engineered soybean MON89788 a kind of, Yi Jiyi
Kind can the method and kit quick, easy, specifically that detect genetically engineered soybean MON89788.
The present invention is realized especially by following technical scheme:
According to the nucleotide sequence of the specificity of transformant of genetically engineered soybean MON89788, genetically engineered soybean is designed
The RPA detection primer of MON89788, the primer include forward primer F and reverse primer R, and nucleotide sequence is specific as follows
It is shown:
Forward primer F:CTTCCTTTTGGGCTTTTTTGTTTCCCGCTCT (SEQ ID NO:1)；
Reverse primer R:GAGCTTGATGGGGATCAGATTGTCGTTTCC
(SEQ ID NO:2).
Application of the above-mentioned primer in RPA detection genetically engineered soybean MON89788 germ plasm resource.
The application on the other hand provide RPA detection genetically engineered soybean MON89788 germ plasm resource probe, the probe with
Above-mentioned primer is used cooperatively, the probe nucleotide sequence are as follows:
P:CGCTTCAATCGTGGTTATCAAGCTCCAAACAC [FAM-dT] [THF] A [BHQ1-dT]
AGTTTAAACTGAAGGC-C3Spacer (SEQ ID NO:3).
A kind of RPA detection kit of genetically engineered soybean MON89788, including above-mentioned forward primer F, reverse primer R and spy
Needle P.
The RPA detection kit specifically includes following components:
1) detection primer and probe solution: forward primer F (10 μm of ol/L), reverse primer R (10 μm of ol/L), probe P (10
μmol/L)；
2) Buffer A: include 50mmol/L Tris-HCl, 8.4 pH, 80mmol/L KAc, 2mmol/L DTT,
3mmol/L ATP, 200 μm of ol/L dNTPs, 20mmol/L C4H10N3O5P, 100ng/ μ L creatine kinase, 5%
20mol/L PEG, 600ng/ μ L SSB, 125ng/ μ L uvsX, 25ng/ μ L uvxY, 30ng/ μ L Bsu and 96ng/ μ L Exo
Ⅲ；
3) BufferB:10mmol/L MgAc；
4) positive control, genetically engineered soybean MON89788 powder (content 0.1%).
On the basis of mentioned reagent box, another aspect of the present invention discloses the RPA of genetically engineered soybean MON89788 a kind of
Detection method, specifically includes the following steps:
1) genomic DNA of sample to be tested is extracted；
2) RPA for preparing sample to be tested detects reaction system: 2 μ L of template DNA, inspection being added in 200 μ L PCR reaction tubes
Survey each 1 μ L of positive and negative primer solution, 0.5 μ L of probe solution, 23.5 μ L of BufferA solution, 2 μ L of BufferB solution, total volume 30
μL；
3) operation RPA amplification judges with result: PCR reaction tube being placed in fluorescence detector, 37~42 DEG C of incubations
20min analyzes amplification according to the presence or absence of fluorescence curve amplification in real time.
Beneficial effects of the present invention:
The present invention is based on recombinase polymeric enzymatic amplification technologies to detect genetically engineered soybean MON89788 germ plasm resource, passes through experiment
It proves, the RPA detection primer and probe combinations of genetically engineered soybean MON89788 provided by the invention, kit and detection method have
Have the advantages that rapidly and efficiently, easy to operate, high specificity.
Detailed description of the invention
Fig. 1 is the result figure that specific detection is carried out in the embodiment of the present invention 4；
Fig. 2 is the result figure that sensitivity technique is carried out in the embodiment of the present invention 5.
Specific embodiment
Below in conjunction with specific embodiment of the present invention, technical solution of the present invention is clearly and completely described, is shown
So, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based on the reality in the present invention
Example is applied, every other embodiment obtained by those of ordinary skill in the art without making creative efforts all belongs to
In the scope of protection of the invention.
The design and screening of embodiment 1 genetically engineered soybean MON89788RPA detection primer and probe
The target area that MON89788RPA is detected in the present invention is the specificity of transformant sequence of MON89788, i.e. external source
The connection region sequence of Insert Fragment and soybean genome, forward and reverse primer therein are located at the two sides of tie point, and probe
Then cover join domain.
(1) design of primers
5 primers, primer length 30-32bp are respectively designed in the two sides of tie point respectively, the present invention screens primer sequence
It is as follows:
F1:GTTTTCCATACTTGATTTGTCCCTCTTGGCT；
F2:TCTAAGTTTGAGCTCGTTACTGCTGCCCCAC；
F3:CCTCGAAACTTGTTCCTGCTCCACTCTTCCT；
F4:TTGTTCCTGCTCCACTCTTCCTTTTGGGCTT；
F5:CTTCCTTTTGGGCTTTTTTGTTTCCCGCTCT；
R1:TACCCTGACCTTGTTGAGGCTTTGGACTGA；
R2:GAATTAGCTTCCACTCGAGCAGGACCTGCA；
R3:GAAGCTTGATAACGCGGCCGCTCTAGCTAGA；
R4:GAGCTTGATGGGGATCAGATTGTCGTTTCC；
R5:GATAACGCGGCCGCTCTAGCTAGAGCTTGAT。
(2) probe designs
Design principle: probe sequence covers join domain, and the length of probe is 46-52bp, in interphase away from 1-5 base
Thymidine (T) on respectively mark fluorescent group, 5 ' end T on mark fluorophor (such as FAM), 3 ' end T on mark it is sudden
Go out group (such as BHQ1), designs abasic site analog in the middle position of two fluorophors, such as tetrahydrofuran (THF),
It is marked in 3 ' ends of probe with C3, it is prevented to extend amplification.As probe and target area pairing, exonuclease (exo
III it) identifies the site THF and cuts probe, separate fluorophor and quencher, to generate fluorescence.
P:CGCTTCAATCGTGGTTATCAAGCTCCAAACAC[FAM-dT][THF]A[BHQ1-dT]
AGTTTAAACTGAAGGC-C3Spacer。
(3) screening strategy of primer and probe
It selects a forward primer and all reverse primers to match, carries out RPA amplification respectively in connection with probe, select expansion
The optimal reverse primer of synergy fruit is now expanded using optimal reverse primer and other forward primer pairings, to select
Optimal forward primer is selected out, best primer pair is F5/R4 in the present invention.
The optimization of 2 RPA reaction system of embodiment
(1) determination of temperature is expanded
Under the other amplification condition unanimous circumstances of RPA, RPA amplification is carried out in 30-45 DEG C of temperature range, according to not
The power of synthermal lower fluorescence signal intensity determines the best amplification temperature of RPA, final to determine that best amplification temperature is 39 DEG C.
(2) concentration of primer and probe determines
Primer is 10 μm of ol/L as the compound concentration of probe, and the other amplification conditions of RPA are consistent, most at 39 DEG C
It expands, various concentration primer is expanded with probe combinations respectively, according to fluorescence signal under various combination at a temperature of good amplification
The power of intensity determines the best primer and concentration and probe concentration of RPA, final to determine that optium concentration is, in the RPA that total volume is 30 μ L
In amplification system, each 1 μ L of positive and negative primer solution, 0.5 μ L of probe solution are detected.
The foundation of 3 kit of embodiment and its detection method
The RPA detection kit of genetically engineered soybean MON89788 is prepared according to the following formula, the specification of each kit is 50
Secondary response:
(1) detection primer and probe solution: synthesis forward primer F, reverse primer R and probe P, by primer and probe dry powder
It is made into the mother liquor that concentration is 10 μm of ol/L respectively with sterile deionized water or ultrapure water, wherein primer sequence is respectively as follows:
Forward primer F:CTTCCTTTTGGGCTTTTTTGTTTCCCGCTCT (SEQ ID NO:1)；
Reverse primer R:GAGCTTGATGGGGATCAGATTGTCGTTTCC
(SEQ ID NO:2)；
Probe P:CGCTTCAATCGTGGTTATCAAGCTCCAAACAC [FAM-dT] [THF] A [BHQ1-dT]
AGTTTAAACTGAAGGC-C3Spacer (SEQ ID NO:3).
(2) BufferA (1.5mL): including 50mmol/L Tris-HCl, 8.4 pH, 80mmol/L KAc, 2mmol/L
DTT, 3mmol/L ATP, 200 μm of ol/L dNTPs, 20mmol/L C4H10N3O5P, 100ng/ μ L creatine kinase,
5%20mol/L PEG, 600ng/ μ L SSB, 125ng/ μ L uvsX, 25ng/ μ L uvxY, 30ng/ μ L Bsu and 96ng/ μ L
ExoⅢ。
(3) Buffer B (120 μ L): 10mmol/L MgAc.
It further include positive control in kit: genetically engineered soybean MON89788 powder (content 0.1%).
Sample to be tested is detected by the following method with mentioned reagent box:
(1) extract the genomic DNA of sample to be tested: traditional DNA extraction method is other with effect commercial kit, extracts
The genomic DNA of sample to be tested is diluted to 25ng/ μ L；
(2) RPA for preparing sample to be tested detects reaction system: using the reaction system of 30 μ L, in 200 μ L PCR reaction tubes
2 μ L of template DNA, each 1 μ L of the positive and negative primer solution of detection, 0.5 μ L of probe solution, 23.5 μ L of Buffer solution A, Buffer is added
2 μ L of B solution
(3) operation RPA amplification judges with result: PCR reaction tube is placed in fluorescence detector, 37~42 DEG C of incubations 30~
60min analyzes amplification according to the presence or absence of fluorescence curve amplification in real time, is if having typical fluorescent amplification curve to occur
The positive is negative if without amplification curve.
The specificity of 4 kit of embodiment and detection method
The specificity of kit described in embodiment 3 and its detection method is tested, test sample both includes turning base
Because of soybean sample, also includes some common transgenic corns, rice, cotton, rape sample, be respectively as follows:
(1) S1: genetically engineered soybean MON89788 (content 1%)；
(2) S2: genetically engineered soybean mixing sample (containing MON89788, GST40-3-2, A5547-127, A2704-12,
356043,1%) content of every kind of transgene component is；
(3) S3: genetically engineered soybean mixing sample (containing MON89788, CV127, MON87701, MON87708,
1%) MON87769, the content of every kind of transgene component are；
(4) S4: (containing MON87705, FG72, the content of every kind of transgene component is genetically engineered soybean mixing sample
1%)；
(5) S5: genetically engineered soybean mixing sample (containing GST40-3-2, A5547-127, A2704-12,356043, every kind
1%) content of transgene component is；
(6) S6: (containing CV127, MON87701, MON87708, MON87769, every kind turns genetically engineered soybean mixing sample
1%) content of gene element is
(7) S7: (containing MON87705, FG72, the content of every kind of transgene component is genetically engineered soybean mixing sample
1%)；
(8) S8: (containing NK603, GA21, TC1507, T25, every kind of transgene component contains transgenic corns mixing sample
1%) amount is；
(9) S9: transgenic corns mixing sample (contains every kind of transgene component of Bt11, Bt176, MON810, MON863
1%) content is
(10) S10: (containing M12, KF-8, KF-2, G6H1, every kind of transgene component contains transgenic paddy rice mixing sample
1%) amount is；
(11) S11: transgenic paddy rice mixing sample (contains TT51-1, KF-6, KMD-1, the content of every kind of transgene component
For 1%)；
(12) S12: transgene cotton mixing sample (containing MON1445, MON531, MON15985, MON88913,
1%) GHB614, the content of every kind of transgene component are；
(13) S13: transgene rape mixing sample (contains MS1, MS8, RF1, RF2, RF3, T45, every kind of transgene component
Content be 1%)
(14) S14: Non-transgenic soybean control；
(15) S15: non-transgenic crop compares (containing non-transgenic corn, rice, cotton, rape)；
(16) S16: ultrapure water blank control.
As a result as shown in Figure 1, only there is curve amplification in the present embodiment in the sample containing genetically engineered soybean MON89788
Curve shows that kit of the present invention and detection method have specificity well to genetically engineered soybean MON89788.
The sensitivity of 5 kit of embodiment and detection method
The sensitivity of kit described in embodiment 3 and its detection method is tested, test sample is that difference turns base
Because of the genetically engineered soybean MON89788 sample of content, it is respectively as follows:
(1) S1: genetically engineered soybean MON89788 (content 1%)；
(2) S2: genetically engineered soybean MON89788 (content 0.1%)；
(3) S3: genetically engineered soybean MON89788 (content 0.05%)；
(4) S4: Non-transgenic soybean control；
(5) S5: ultrapure water blank control.
As a result as shown in Fig. 2, in the present embodiment containing genetically engineered soybean MON89788 content be respectively 1%, 0.1% and
There is good amplification curve in 0.05% sample, shows kit of the present invention and detection method to genetically engineered soybean
MON89788 have good sensitivity, at least up to 0.05%.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
Understand without departing from the principles and spirit of the present invention can to these examples carry out it is a variety of variation, modification, replacement and
Modification, the scope of the present invention is defined by the appended.
Sequence table
<110>Zhejiang Academy of Agricultural Science
<120>a kind of primer, probe and kit and method for detecting genetically engineered soybean MON89788
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 31
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 1
cttccttttg ggcttttttg tttcccgctc t 31
<210> 2
<211> 30
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
gagcttgatg gggatcagat tgtcgtttcc 30
<210> 3
<211> 49
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 3
cgcttcaatc gtggttatca agctccaaac acaagtttaa actgaaggc 49
Claims (9)
1. a kind of RPA detection primer of genetically engineered soybean MON89788, which is characterized in that the primer includes forward primer F
With reverse primer R, the nucleotide sequence of the forward primer F is as shown in SEQ ID NO.1, the core of the reverse primer R
Nucleotide sequence is as shown in SEQ ID NO.2.
2. application of the RPA detection primer described in claim 1 in detection genetically engineered soybean MON89788 germ plasm resource.
3. a kind of probe of RPA detection genetically engineered soybean MON89788 germ plasm resource, which is characterized in that the probe and right
It is required that primer described in 1 is used cooperatively, the probe nucleotide sequence is as shown in SEQ ID NO.3.
4. a kind of RPA detection kit of genetically engineered soybean MON89788, which is characterized in that including it is described in claim 1 just
To primers F and reverse primer R.
5. the RPA detection kit of genetically engineered soybean MON89788 according to claim 4 a kind of, which is characterized in that packet
Include probe as claimed in claim 3.
6. a kind of RPA detection method of genetically engineered soybean MON89788, which comprises the following steps:
1) genomic DNA of sample to be tested is extracted；
2) RPA for preparing sample to be tested detects reaction system: 2 μ L of template DNA, detection being added in 200 μ L PCR reaction tubes just
Each 1 μ L of anti-primer solution, 0.5 μ L of probe solution, 23.5 μ L of BufferA solution, 2 μ L of BufferB solution, total volume are 30 μ L；
3) operation RPA amplification judges with result: PCR reaction tube being placed in fluorescence detector, 37~42 DEG C of incubation 20min, root
According to the presence or absence of fluorescence curve amplification, amplification is analyzed in real time.
7. detection method according to claim 6, which is characterized in that the concentration of the positive and negative primer and probe solution is
10μmol/L。
8. detection method according to claim 6, which is characterized in that the Buffer A includes: 50mmol/L
Tris-HCl, pH 8.4,80mmol/L KAc, 2mmol/L DTT, 3mmol/L ATP, 200 μm of ol/L dNTPs, 20mmol/L
C4H10N3O5P, 100ng/ μ L creatine kinase, 5%20mol/L PEG, 600ng/ μ L SSB, 125ng/ μ L uvsX,
25ng/ μ L uvxY, 30ng/ μ L Bsu and 96ng/ μ L Exo III.
9. detection method according to claim 6, which is characterized in that the Buffer B is 10mmol/L MgAc.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811222863.8A CN109136338A (en) | 2018-10-19 | 2018-10-19 | A kind of primer, probe and kit and method detecting genetically engineered soybean MON89788 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811222863.8A CN109136338A (en) | 2018-10-19 | 2018-10-19 | A kind of primer, probe and kit and method detecting genetically engineered soybean MON89788 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109136338A true CN109136338A (en) | 2019-01-04 |
Family
ID=64809006
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811222863.8A Pending CN109136338A (en) | 2018-10-19 | 2018-10-19 | A kind of primer, probe and kit and method detecting genetically engineered soybean MON89788 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109136338A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110512023A (en) * | 2019-09-04 | 2019-11-29 | 中国农业科学院油料作物研究所 | Method based on the identification soybean transformants MON89788 genotype that insertion point genome sequence is established |
| CN112359127A (en) * | 2020-11-13 | 2021-02-12 | 浙江省农业科学院 | RPA primer and probe combination for detecting transgenic soybean SHZD32-1 and on-site rapid detection method |
| CN115992284A (en) * | 2022-09-16 | 2023-04-21 | 上海市农业科学院 | Kit and method for rapidly detecting transgenic product based on RPA-CRISPR-Cas12a system |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104513861A (en) * | 2015-01-14 | 2015-04-15 | 上海速芯生物科技有限公司 | LAMP test primer set, detection kit and detection method for transgenic soybean MON89788 and derived varieties of transgenic soybean MON89788 |
| CN104726582A (en) * | 2015-03-17 | 2015-06-24 | 苏州华麦生物科技有限公司 | Probe method based constant temperature detection primer group, detection kit and detection method of genetically modified soybeans MON89788 |
| CN106755488A (en) * | 2017-01-22 | 2017-05-31 | 中国农业科学院生物技术研究所 | Transgenic corn BT 11 strain specificity is identified using RPA technologies |
-
2018
- 2018-10-19 CN CN201811222863.8A patent/CN109136338A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104513861A (en) * | 2015-01-14 | 2015-04-15 | 上海速芯生物科技有限公司 | LAMP test primer set, detection kit and detection method for transgenic soybean MON89788 and derived varieties of transgenic soybean MON89788 |
| CN104726582A (en) * | 2015-03-17 | 2015-06-24 | 苏州华麦生物科技有限公司 | Probe method based constant temperature detection primer group, detection kit and detection method of genetically modified soybeans MON89788 |
| CN106755488A (en) * | 2017-01-22 | 2017-05-31 | 中国农业科学院生物技术研究所 | Transgenic corn BT 11 strain specificity is identified using RPA technologies |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110512023A (en) * | 2019-09-04 | 2019-11-29 | 中国农业科学院油料作物研究所 | Method based on the identification soybean transformants MON89788 genotype that insertion point genome sequence is established |
| CN112359127A (en) * | 2020-11-13 | 2021-02-12 | 浙江省农业科学院 | RPA primer and probe combination for detecting transgenic soybean SHZD32-1 and on-site rapid detection method |
| CN115992284A (en) * | 2022-09-16 | 2023-04-21 | 上海市农业科学院 | Kit and method for rapidly detecting transgenic product based on RPA-CRISPR-Cas12a system |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109337959A (en) | A kind of RPA detection primer of transgenic corns MON810 and probe combinations, kit and detection method | |
| CN109136338A (en) | A kind of primer, probe and kit and method detecting genetically engineered soybean MON89788 | |
| CN106399490B (en) | LAMP primer group for detecting phytoplasma and kit and application thereof | |
| CN109136341A (en) | A kind of primer, probe and kit and method detecting genetically engineered soybean A2704-12 | |
| KR20190092821A (en) | Primer set for detection of Foot-and-Mouth Disease virus and use thereof | |
| CN103924000B (en) | LAMP detecting primer group, kit and detecting method for cry3A gene in transgenic plant | |
| CN109055501A (en) | A kind of primer, probe and kit and method detecting genetically engineered soybean GST40-3-2 | |
| CN109136340A (en) | A kind of primer, probe and kit and method detecting transgene rape Oxy-235 | |
| Nguyen et al. | Critical role for uricase and xanthine dehydrogenase in soybean nitrogen fixation and nodule development | |
| CN104513861B (en) | LAMP detection primer group, detection kit and the detection method of genetically engineered soybean MON89788 and its derived varieties | |
| CN110241245A (en) | Detect KASP primer and its application of cucumber bacterial angular leaf spot gene | |
| CN109182585A (en) | A kind of primer, probe and kit and method detecting transgenic corns TC1507 | |
| CN109161585A (en) | A kind of primer, probe and kit and method detecting transgenic corns MIR604 | |
| CN107815490A (en) | A kind of tobacco root-knot pathogeny detecting reagent box and its application method based on loop-mediated isothermal amplification technique | |
| CN106755545A (en) | The LAMP detection primer group of pat gene, kit and detection method in genetically modified crops | |
| CN116516059A (en) | Method and kit for visually detecting phoma sojae atricolor | |
| CN105063207B (en) | Genetically engineered soybean MON87705 LAMP detection primer group, kit and detection method | |
| CN109055592A (en) | A kind of anti-ring spot virus papaya YK16-0-1 of transgenosis and its efficient qualitative, quantitative identification method of spin-off | |
| CN109234434A (en) | A kind of primer, probe and kit and method detecting transgenic paddy rice G6H1 | |
| CN109280717A (en) | A kind of RPA detection primer of transgene rape T45 and probe combinations, kit and detection method | |
| CN116083541A (en) | Method for enriching low-abundance mononucleotide variant | |
| CN109136339A (en) | A kind of primer, probe and kit and method detecting transgenic paddy rice TT51-1 | |
| CN110564822B (en) | LAMP technology-based transgenic corn Bt 176-related gene detection method and kit | |
| CN106399540A (en) | Corn detection method based on loop-mediated isothermal amplification extinction technology | |
| CN104726580B (en) | A kind of transgenic corns MON810 constant temperature probe method detection primer group, detection kit and detection method |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190104 |