CN113881758B - LAMP constant temperature amplification method for detecting coconut ingredients in vegetable protein beverage - Google Patents
LAMP constant temperature amplification method for detecting coconut ingredients in vegetable protein beverage Download PDFInfo
- Publication number
- CN113881758B CN113881758B CN202111332830.0A CN202111332830A CN113881758B CN 113881758 B CN113881758 B CN 113881758B CN 202111332830 A CN202111332830 A CN 202111332830A CN 113881758 B CN113881758 B CN 113881758B
- Authority
- CN
- China
- Prior art keywords
- coconut
- protein beverage
- amplification
- lamp
- vegetable protein
- 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.)
- Active
Links
Images
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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (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 invention provides an LAMP (loop-mediated isothermal amplification) method for detecting coconut components in vegetable protein beverage, which comprises the following steps: 1. preparing a plant protein beverage DNA sample and LAMP constant temperature amplification specific primers; after the DNA sample of the vegetable protein beverage is measured by an ultramicro spectrophotometer, the A260/A280 of the DNA sample is between 1.7 and 2.0, and the DNA concentration is more than 10 ng/mu L; the LAMP constant temperature amplification specific primer is synthesized by selecting a coconut specific gene sequence, and the coconut gene adopts a coconut 3a (Hd 3 a) gene; the position of the specific gene sequence in the coconut gene is 451-1300bp; the LAMP constant temperature amplification specific primer is as follows: an outer primer F3/B3, an inner primer FIB/BIP and a loop primer LB; 2. and (3) performing LAMP isothermal amplification on the plant protein beverage DNA sample by adopting an LAMP isothermal amplification reaction system, and detecting a fluorescent signal to obtain a detection result. The method has high sensitivity and high specificity for identifying the coconut components, and can be used for quickly detecting the coconut components in the vegetable protein beverage.
Description
Technical Field
The invention belongs to the technical field of molecular biology, and particularly relates to an LAMP constant temperature amplification method for detecting coconut ingredients in vegetable protein beverages.
Background
Coconut is the fruit of coconut tree of palmaceae, coconut juice and coconut flesh contain protein, sugar, grease, vitamins, calcium, phosphorus, iron and other trace elements and mineral substances, and are popular with consumers. In recent years, vegetable protein beverages (such as coconut juice) using coconut as raw material have been rapidly developed by virtue of their "nature, green, nutritional, health" and other advantages, and are currently in the market in a heavier proportion. With the rapid increase of market demand and the increase of raw material prices, some production enterprises forge coconut beverages by using low-priced soybeans or other cheap raw materials in order to reduce production costs, and seize the market at low prices. At present, the method for detecting coconut ingredients of vegetable protein beverage has no relevant detection standard, only common PCR detection method is seen in relevant documents, the vegetable protein beverage is widely distributed in the market, the common PCR detection technology is more complicated in flow, the negative and positive are judged by electrophoresis results, the whole detection process cannot be monitored in real time, in addition, the requirement on operators is higher, and the method is difficult to popularize and apply in a large number in the vast base.
LAMP detection technology, called Loop-mediated isothermal amplification (LAMP), was the first isothermal Nucleic acid amplification technology disclosed by Japanese scholars in Nucleic Acids Res for gene diagnosis, and its main principle is to use Bst DNA polymerase with strand displacement specificity and 4-6 primers capable of specifically recognizing multiple specific regions on the target sequence to start the cycle strand displacement reaction, thereby realizing continuous and rapid reaction under isothermal condition. The technology has the advantages of strong specificity, high sensitivity, low detection cost, low requirements on required equipment and personnel, simple operation, short reaction time and the like, and is suitable for rapid detection of large-batch samples on site.
Therefore, it is necessary to establish a set of LAMP constant temperature amplification method for coconut ingredients of vegetable protein beverage, which has good specificity and high sensitivity and is suitable for being applied in the basement level and on-site security, so as to provide an efficient detection method for resisting the problem of adulteration of coconut protein beverage and provide a new technical support for guaranteeing the food quality and safety of the beverage industry.
Disclosure of Invention
The invention aims to provide an LAMP constant temperature amplification method for detecting the coconut ingredients in the vegetable protein beverage, which has high sensitivity and specificity on the identification of the coconut ingredients and can be used for quickly detecting the coconut ingredients in the vegetable protein beverage.
The purpose of the invention can be realized by the following technical scheme:
an LAMP constant temperature amplification method for detecting coconut ingredients in vegetable protein beverage comprises the following steps:
step 1, preparing a plant protein beverage DNA sample and preparing LAMP constant temperature amplification specific primers for performing LAMP constant temperature amplification on the plant protein beverage DNA sample;
after the DNA sample of the vegetable protein beverage is measured by an ultramicro spectrophotometer, the A260/A280 of the DNA sample is between 1.7 and 2.0, the DNA concentration is more than 10 ng/mu L, and the DNA sample is stored at the temperature of minus 20 ℃ for later use;
the LAMP constant-temperature amplification specific primer is obtained by selecting a coconut specific gene sequence for synthesis, and the coconut gene adopts the following steps: the coconut 3a (Hd 3 a) gene with accession number MH092984.1; the position of the specific gene sequence in the coconut gene is 451-1300bp;
the LAMP constant temperature amplification specific primer is as follows:
an outer primer F3:5'-CTCCAAGTCCAAGTGACC-3'
An outer primer B3:5'-GTGGAGCTAGTTTTGTTCC-3'
Inner primer FIB:5'-CATGGCTTCCCGATGGGTACCTCAGGGAGTATTTGCACTG-3'
The inner primer BIP:5'-GCTATATATTGGGCCGGACTTCGCAATCTCTTCTCTTCCTTGT-3'
The loop primer LB:5'-ACTCTTCAGCGGGTGCTTAC-3'
Step 2, performing LAMP isothermal amplification on the plant protein beverage DNA sample by adopting an LAMP isothermal amplification reaction system, and detecting a fluorescent signal of an amplification product to obtain a detection result; wherein the LAMP isothermal amplification reaction system comprises the LAMP isothermal amplification specific primer.
Optimizing the scheme, when the vegetable protein beverage in the step 1 is solid or liquid containing solid, the solid vegetable protein beverage is mixed with warm water according to the specification of the solid vegetable protein beverage, and the mixture is uniformly stirred to obtain a liquid vegetable protein beverage sample to be extracted; fully grinding the solid matter in the liquid vegetable protein beverage containing the solid matter by using a grinder to obtain a liquid vegetable protein beverage sample to be extracted; and (3) carrying out DNA extraction on the plant protein beverage sample to be extracted to obtain the plant protein beverage DNA sample in the step (1).
The LAMP constant-temperature amplification reaction system is 25 mu L, and comprises 0.4 mu L of each of an inner primer FIP (100 mu mol/L) and an inner primer BIP (100 mu mol/L), 0.05 mu L of each of an outer primer F3 (100 mu mol/L) and an outer primer B3 (100 mu mol/L) and 0.2 mu L of a loop primer LB (100 mu mol/L); the other reagent components are respectively as follows: 10 Xthermo pol Buffer 2.5. Mu.L, mgSO 4 1. Mu.L (100 mmol/L), 2.5. Mu.L dNTPs (10 mmol/L), 6. Mu.L betaine (5 mol/L), 1. Mu.L Bst enzyme (8U/. Mu.L), 0.5. Mu.L dye (250X), 2. Mu.L DNA template, ddH 2 O 8.4μL。
In the LAMP isothermal amplification reaction system, the LAMP isothermal amplification conditions of an isothermal amplification instrument are as follows: the amplification temperature was 63 ℃ and 45 cycles were performed, with an amplification time of 60s per cycle; the amplification conditions of the real-time fluorescent PCR instrument are as follows: the first step is amplification for 30s at 63 ℃, the second step is amplification for 15s at 63 ℃ and 45 cycles, and the third step is amplification for 45s at 63 ℃ and 45 cycles.
The invention has the following prominent substantive features and remarkable progress:
1. the invention provides a specific gene sequence for identifying coconut ingredients, which has high specificity, can be used for qualitative detection of the coconut ingredients and has commercial development value. The applicant carries out specificity detection on 45 plant and animal material DNA samples by using the LAMP constant temperature amplification specific primer and the LAMP constant temperature amplification reaction system designed by the specific gene sequence, and the result shows that the specific gene sequence has high specificity and can be used as a specific marker for qualitative detection of coconut components, wherein the amplification of the specific gene sequence is positive for coconut DNA samples, and the amplification of the specific gene sequence is negative for DNA samples of other various species. In addition, the applicant also carries out sensitivity detection and stability detection on the method, and the result shows that the method has the characteristics of high sensitivity and stability.
2. The LAMP isothermal amplification specific primer capable of specifically identifying the coconut ingredients is provided in the LAMP isothermal amplification method for detecting the vegetable protein beverage coconut ingredients, the specificity is good, the specific detection can be realized on the coconut-derived ingredients, and other ingredients except the coconut cannot be specifically amplified; in addition, the invention also provides an LAMP isothermal amplification reaction system, which can realize whether the sample contains coconut components within 1 hour and has the characteristics of high efficiency, rapidness, convenience and convenience in detection.
3. The LAMP constant-temperature amplification specific primer can be used for quickly detecting whether the coconut ingredients exist in the vegetable protein beverage or not at high flux, and the required instruments and equipment are low in cost, simple to operate, high in efficiency and low in requirement on inspection personnel, so that the LAMP constant-temperature amplification specific primer is suitable for quick detection of vast primary and on-site guarantees.
4. The LAMP constant-temperature amplification method for detecting the coconut components of the vegetable protein beverage is suitable for detecting the coconut components of various vegetable protein beverages, including solid beverages, liquid beverages, semi-solid beverages and the like, can be popularized and applied in a large quantity, provides technical support for guaranteeing the quality of products, protecting rights and interests of consumers, maintaining normal beverage industry order, and provides technical support for food market supervision and management departments.
Drawings
FIG. 1 is a LAMP amplification map of 20 coconut components as vegetable protein beverage by the method of the present invention.
FIG. 2 is a LAMP-specific amplification map for verifying the detection of coconut components using the method of the present invention.
FIG. 3 is a LAMP sensitivity amplification map for verifying the coconut component detection using the method of the present invention.
FIG. 4 is a LAMP stability amplification map demonstrating coconut component detection using the method of the present invention.
Detailed Description
The invention will be further described with reference to the accompanying drawings.
Example 1
An LAMP constant temperature amplification method for detecting coconut ingredients in vegetable protein beverage comprises the following steps:
step 1, preparing a plant protein beverage DNA sample and preparing LAMP constant temperature amplification specific primers for performing LAMP constant temperature amplification on the plant protein beverage DNA sample;
in the step, when the plant protein beverage to be detected is solid or liquid containing solid, the solid plant protein beverage is mixed with warm water according to the specification of the solid plant protein beverage, and the mixture is stirred uniformly to obtain a liquid plant protein beverage sample to be extracted; fully grinding the solid matter in the liquid vegetable protein beverage containing the solid matter by using a grinder to obtain a liquid vegetable protein beverage sample to be extracted;
extracting plant protein beverage DNA by using a magnetic bead method (Liang Meidan, xiaojian, chen Jie, huang Zhishen, laojiaqian. Food safety quality detection academic newspaper, 2021, 12 (13): 5363-5368), extracting the DNA of a sample to be extracted of the plant protein beverage, wherein the extraction method is detailed in literature data to obtain the plant protein beverage DNA sample; after the DNA sample of the vegetable protein beverage is measured by an ultramicro spectrophotometer, the A260/A280 of the DNA sample is between 1.7 and 2.0, the DNA concentration is more than 10 ng/mu L, and the DNA sample is stored at the temperature of minus 20 ℃ for later use;
the LAMP constant-temperature amplification specific primer is obtained by selecting a coconut specific gene sequence for design and synthesis. The method comprises the following steps: coconut (Cocos nucifera) 3a (Hd 3 a) gene (accession number: MH 092984.1) is downloaded from NCBI website (https:// www.ncbi.nlm.nih.gov /), specific gene sequence is selected, the position is 451-1300bp, a group of LAMP constant temperature amplification primers is designed by using Primer Explorer Version 4 (http:// Primer Explorer. Jp/e/v4_ manual/index. Html):
an outer primer F3:5'-CTCCAAGTCCAAGTGACC-3'
An outer primer B3:5'-GTGGAGCTAGTTTTGTTCC-3'
Inner primer FIB:5'-CATGGCTTCCCGATGGGTACCTCAGGGAGTATTTGCACTG-3'
The inner primer BIP:5'-GCTATATATTGGGCCGGACTTCGCAATCTCTTCTCTTCCTTGT-3'
The loop primer LB:5'-ACTCTTCAGCGGGTGCTTAC-3'
Step 2, performing LAMP isothermal amplification on the plant protein beverage DNA sample by adopting an LAMP isothermal amplification reaction system, and detecting a fluorescent signal of an amplification product to obtain a detection result; the LAMP isothermal amplification reaction system comprises the LAMP isothermal amplification specific primer, and the specific details are shown in Table 1.
The reaction program of the LAMP constant temperature amplification reaction system is as follows:
the LAMP isothermal amplification conditions of the isothermal amplification instrument are as follows: the amplification temperature is 63 ℃, 45 cycles are carried out, the amplification time of each cycle is 60s, and the total amplification time is 45min;
the amplification conditions of the real-time fluorescent PCR instrument are as follows: the amplification is carried out for 30s at 63 ℃, for 15s and 45 cycles at 63 ℃ in the first step, and for 45s and 45 cycles at 63 ℃ in the third step, wherein the total amplification time is about 50min.
In this example, the coconut component was detected on 20 DNA samples of vegetable protein beverage, and the names of the 20 DNA samples of vegetable protein beverage and the detection results of the coconut component are shown in FIG. 1 and Table 2.
As can be seen from FIG. 1 and Table 2, in 6 vegetable protein beverages containing coconut components, the test results of the present invention are completely consistent with the components indicated in the product specification; the rest 12 samples contain walnut, sesame, hazelnut, almond, peanut and soybean components in a marked way, and the detection results of the invention are all negative results.
And (4) conclusion: 6 samples claim to contain the vegetable protein beverage of coconut ingredient, the invention detects the coconut ingredient as positive, 12 samples claim not to contain the vegetable protein beverage of coconut ingredient, the invention detects the coconut ingredient as negative, the detection result is 100% accord with. Meanwhile, the detection method of the invention can be used for detecting whether coconut ingredients exist in the vegetable protein beverage in large batch.
In conclusion, the LAMP isothermal amplification specific primer and the LAMP isothermal amplification reaction system for the coconut ingredients designed by the invention have the advantages of good specificity and high sensitivity, can quickly realize the qualitative detection of the coconut ingredients in the vegetable protein beverage, do not need to depend on high-end expensive instruments and equipment, have low detection cost and lower requirements on professional technicians, are suitable for the quick detection of basic and on-site guarantees, and have good application prospects. In addition, the LAMP constant-temperature amplification specific primer can be further prepared into an LAMP detection kit for the coconut components, is applied to detection kits for the coconut components in food and other samples, and has wide application range and strong practicability.
Example 2
In this embodiment, the specificity of the LAMP isothermal amplification reaction system and the LAMP isothermal amplification specific primers of the present invention is detected, specifically: the LAMP isothermal amplification reaction system of example 1 was used to detect DNA samples of 45 plant and animal materials tested in this example, and the fluorescence signal of the amplified product was detected. The results are shown in FIG. 2 and Table 3.
As can be seen from FIG. 2 and Table 3, the present invention has positive amplification and obvious sigmoid amplification curve for coconut DNA samples, and negative amplification and no obvious sigmoid amplification curve for DNA samples of other various species.
And (4) conclusion: the detection method of the invention has good species specificity.
Example 3
The embodiment is to detect the LAMP isothermal amplification reaction system and the sensitivity of LAMP isothermal amplification specific primers, and specifically comprises the following steps: the method comprises the following steps of preparing coconut mixed samples with different mass fractions by taking mung beans as a matrix according to a proportion, wherein the mass fractions of coconut DNA in the coconut mixed samples comprise: 50%, 10%, 5%, 1%, 0.1%, 0.01%, wherein 0.01% of coconut mixed sample is obtained by diluting 10 times of coconut mixed sample DNA with the mass fraction of 0.1%, performing LAMP isothermal amplification according to the LAMP isothermal amplification reaction system and conditions of example 1, and repeating the experiment for 6 times.
As a result: the coconut LAMP fluorescent amplification samples with the mass fractions of 50%, 10%, 5%, 1% and 0.1% all have obvious S-shaped amplification curves, and the coconut mixed sample with the mass fraction of 0.01% does not have an S-shaped amplification curve, which indicates that the sensitivity of the method is 0.1%, and the amplification map is shown in figure 3.
Example 4
The embodiment is to detect the stability of the LAMP isothermal amplification reaction system and the LAMP isothermal amplification specific primers, and specifically comprises the following steps: performing LAMP isothermal amplification on the coconut mixed sample DNA with the mass fraction of 0.1% in the example 3 according to the LAMP isothermal amplification reaction system and conditions in the example 1, repeating for 10 times, and as a result, the coconut mixed sample with the mass fraction of 0.1% can amplify an obvious S-shaped curve, which proves that the method has good stability, and the amplification map is shown in figure 4.
Sequence listing
<110> Guangzhou city food inspection institute
<120> LAMP rapid detection method for detecting coconut ingredients in vegetable protein beverage
<160> 5
<170> SIPOSequenceListing 1.0
<210> 1
<211> 18
<212> DNA
<213> Artificial sequence ()
<400> 1
ctccaagtcc aagtgacc 18
<210> 2
<211> 19
<212> DNA
<213> Artificial sequence ()
<400> 2
gtggagctag ttttgttcc 19
<210> 3
<211> 40
<212> DNA
<213> Artificial sequence ()
<400> 3
catggcttcc cgatgggtac ctcagggagt atttgcactg 40
<210> 4
<211> 43
<212> DNA
<213> Artificial sequence ()
<400> 4
gctatatatt gggccggact tcgcaatctc ttctcttcct tgt 43
<210> 5
<211> 20
<212> DNA
<213> Artificial sequence ()
<400> 5
Claims (4)
1. An LAMP isothermal amplification method for detecting coconut ingredients in vegetable protein beverage is characterized by comprising the following steps:
step 1, preparing a plant protein beverage DNA sample and preparing LAMP constant temperature amplification specific primers for performing LAMP constant temperature amplification on the plant protein beverage DNA sample;
after the DNA sample of the vegetable protein beverage is measured by an ultramicro spectrophotometer, the A260/A280 of the DNA sample is between 1.7 and 2.0, the DNA concentration is more than 10 ng/mu L, and the DNA sample is stored at the temperature of minus 20 ℃ for later use;
the LAMP constant-temperature amplification specific primer is obtained by selecting a coconut specific gene sequence for design and synthesis, wherein the position of the specific gene sequence in a coconut Hd3a gene is 451-1300bp, and the accession number of the coconut Hd3a gene is MH092984.1;
the LAMP constant temperature amplification specific primer is as follows:
an outer primer F3:5'-CTCCAAGTCCAAGTGACC-3'
An outer primer B3:5'-GTGGAGCTAGTTTTGTTCC-3'
Inner primer FIB:
5’-CATGGCTTCCCGATGGGTACCTCAGGGAGTATTTGCACTG-3’
the inner primer BIP:
5’-GCTATATATTGGGCCGGACTTCGCAATCTCTTCTCTTCCTTGT-3’
the loop primer LB:5'-ACTCTTCAGCGGGTGCTTAC-3'
Step 2, performing LAMP isothermal amplification on the plant protein beverage DNA sample by adopting an LAMP isothermal amplification reaction system, and detecting a fluorescent signal of an amplification product to obtain a detection result; the LAMP isothermal amplification reaction system comprises the LAMP isothermal amplification specific primer.
2. The LAMP isothermal amplification method for detecting the vegetable protein beverage coconut ingredients according to claim 1, characterized in that: when the vegetable protein beverage is solid or liquid containing solid in the step 1, the solid vegetable protein beverage is mixed with warm water and stirred uniformly to obtain a liquid vegetable protein beverage sample to be extracted; fully grinding the solid matter in the liquid vegetable protein beverage containing the solid matter by using a grinder to obtain a liquid vegetable protein beverage sample to be extracted; and (3) carrying out DNA extraction on the plant protein beverage sample to be extracted to obtain the plant protein beverage DNA sample in the step (1).
3. The LAMP isothermal amplification method for detecting the vegetable protein beverage coconut ingredients according to claim 1 or 2, characterized in that: the LAMP constant-temperature amplification reaction system is 25 mu L, and comprises 0.4 mu L of each of 100 mu mol/L inner primer FIP and 100 mu mol/L inner primer BIP, 0.05 mu L of each of 100 mu mol/L outer primer F3 and 100 mu mol/L outer primer B3, and 0.2 mu L loop primer LB of 100 mu mol/L; the other reagent components are respectively as follows: 10 × Thermo pol Buffer2.5 μ L, mgSO 4 100 mmol/L1. Mu.L, dNTPs 10 mmol/L2.5. Mu.L, betaine 5 mol/L6. Mu.L, bstase 8U/. Mu.L 1. Mu.L, 250 Xdye 0.5. Mu.L, DNA template 2. Mu.L, ddH 2 O 8.4μL。
4. The LAMP isothermal amplification method for detecting vegetable protein beverage coconut ingredients according to claim 3, characterized in that: in the LAMP isothermal amplification reaction system, the LAMP isothermal amplification conditions of the isothermal amplification instrument are as follows: the amplification temperature was 63 ℃ and 45 cycles were performed, with an amplification time of 60s per cycle; the fluorescent signal amplification conditions of the real-time fluorescent PCR instrument are as follows: the first step is amplification for 30s at 63 ℃, the second step is amplification for 15s at 63 ℃ and 45 cycles, and the third step is amplification for 45s at 63 ℃ and 45 cycles.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111332830.0A CN113881758B (en) | 2021-11-11 | 2021-11-11 | LAMP constant temperature amplification method for detecting coconut ingredients in vegetable protein beverage |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111332830.0A CN113881758B (en) | 2021-11-11 | 2021-11-11 | LAMP constant temperature amplification method for detecting coconut ingredients in vegetable protein beverage |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113881758A CN113881758A (en) | 2022-01-04 |
| CN113881758B true CN113881758B (en) | 2022-11-18 |
Family
ID=79017916
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111332830.0A Active CN113881758B (en) | 2021-11-11 | 2021-11-11 | LAMP constant temperature amplification method for detecting coconut ingredients in vegetable protein beverage |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113881758B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014077775A1 (en) * | 2012-11-16 | 2014-05-22 | Temasek Life Sciences Laboratory Limited | Flowering modification in the palm family |
| CN104502582A (en) * | 2014-12-12 | 2015-04-08 | 深圳市计量质量检测研究院 | Enzyme linked immunosorbent assay kit for detecting coconut protein |
| CN111455093A (en) * | 2020-06-08 | 2020-07-28 | 中国食品药品检定研究院 | Universal primer for identifying plant-derived components and application thereof |
-
2021
- 2021-11-11 CN CN202111332830.0A patent/CN113881758B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014077775A1 (en) * | 2012-11-16 | 2014-05-22 | Temasek Life Sciences Laboratory Limited | Flowering modification in the palm family |
| CN104502582A (en) * | 2014-12-12 | 2015-04-08 | 深圳市计量质量检测研究院 | Enzyme linked immunosorbent assay kit for detecting coconut protein |
| CN111455093A (en) * | 2020-06-08 | 2020-07-28 | 中国食品药品检定研究院 | Universal primer for identifying plant-derived components and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| Jasmin Wrage等.Development of a DNA-Based Detection Method for Cocos Nucifera Using TaqManReal-Time PCR.《Foods》.2020,第9卷(第332期), * |
| 市售椰子汁(植物蛋白饮料)中椰子、大豆、花生源性成分鉴定的分子生物学方法;杨硕等;《基因组学与应用生物学》;20170814(第10期);第288-294页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113881758A (en) | 2022-01-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106811513B (en) | Eucalyptus component real-time fluorescence PCR detection method and kit thereof | |
| CN103397101A (en) | Fluorescently-labeled gene multiplex amplification method for identifying sources of goat, sheep, pig and duck meat simultaneously | |
| CN104293778B (en) | Establishing method of cymbidium microsatellite labels, core fingerprint label database and kit | |
| CN112322766B (en) | Accurate pseudo-ginseng powder quantitative method based on micro-drop digital PCR technology | |
| CN114182030B (en) | LAMP constant-temperature rapid detection method for Burkholderia gladioli | |
| CN113151567B (en) | SSR molecular marker and method for identifying Lepista sordida N006# strain | |
| CN113881758B (en) | LAMP constant temperature amplification method for detecting coconut ingredients in vegetable protein beverage | |
| CN106048029A (en) | LAMP (Loop-mediated Isothermal Amplification) detection primer set, detection kit and detection method of mycoplasma hyopneumonia | |
| CN106701746B (en) | High-throughput malt Purity technology based on Capillary Electrophoresis and SSR marker | |
| CN103525936A (en) | Specific identification for transgenic rice kefeng No.6 strain via adoption of RPA (Recombinase Ploymerase Amplification) technology | |
| CN114182037B (en) | DNA bar code for screening content index of total soluble protein of stropharia rugoso-annulata | |
| CN104630333B (en) | Constant-temperature amplification detection primer, kit and the method for Queensland nut anaphylactogen | |
| CN114231603B (en) | Primer, reagent, identification method and kit for identifying paeonia boita | |
| CN106434976B (en) | Method for identifying true and false of Chinese yam and special primer | |
| CN114150079B (en) | DNA bar code for screening yellow green stropharia rugoso-annulata with high total polysaccharide content | |
| CN104962640A (en) | Method for quickly identifying authenticity of conventional cotton varieties by using SSR (Simple Sequence Repeat) marker | |
| CN110734998A (en) | Primers, method and kit for identifying NFC orange juice and FC orange juice | |
| CN110628939B (en) | Primer probe, method and kit for detecting camellia oleifera derived components | |
| CN110317858B (en) | Method for quantitatively detecting kidney bean components in lotus paste by using dual digital PCR (polymerase chain reaction) | |
| CN113215273B (en) | miRNA specific marker related to meat variety identification and application thereof | |
| CN112941226B (en) | Application of gene spt5 in detection of boletus meiloti derived components | |
| CN118186143B (en) | SSR core primer combination, kit and application for identifying tea tree variety Yinghong Jiu | |
| CN111593133B (en) | Method, composition and kit for identifying four kinds of pollen by multiplex fluorescence PCR | |
| CN107287344A (en) | A kind of kit and detection method for detecting chicken WNT9A gene expression amounts | |
| CN111593134B (en) | Method, composition and kit for identifying four bee pollen by multiple fluorescence PCR |
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 |