CN1597983A - Process of extracting DNA for testing transgene component from sauce - Google Patents
Process of extracting DNA for testing transgene component from sauce Download PDFInfo
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- CN1597983A CN1597983A CN 200410020177 CN200410020177A CN1597983A CN 1597983 A CN1597983 A CN 1597983A CN 200410020177 CN200410020177 CN 200410020177 CN 200410020177 A CN200410020177 A CN 200410020177A CN 1597983 A CN1597983 A CN 1597983A
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Abstract
The invention relates to a method of distilling the DNA from the sauce, which is used in detecting the components of the transgene teatures that adding the pretreatment to the procedure of distilling the DNA to get rid of the caramel pigment in the sauce and to reduce the pressure in the subsequent distilment; increasing the times of distilling the chloroform + isoamyl alcohol and the time of depositing the bromohexadecane trimethylamine to get rid of the protein, polysaccharide and other impurities; resolving the DNA solution into the 30 mul deionized water, getting 15mul for once PCR reaction to increase the density of the DNA cyclostyle in the reaction system. The method is featured by high speed, convenience, strong specificity and high efficiency.
Description
Technical field
Present method relates to the detection technique of transgene component in the deep processing food, especially from being to extract the method that is used for the template DNA that transgene component detects the fermented products such as soy sauce that process of raw material with the soybean.
Background technology
Fast development along with genetic engineering technique, a large amount of is that the genetically modified food that processes raw material enters the commercialization production and selling with genetically modified crops, add up according to internal authority mechanism, kind surplus the whole world reaches 4000 by the genetically modified food of genetically modified crops processing and food ingredient (referring to: U.S.Food and Drug Administration, Office of Food AdditiveSafety.List of Completed Consultations on Bioengineering Food[DB/OL] .http: //www.fda-efsan/biotechnology.2002-10.).
At present still disputable to the security of genetically modified food.The potential risk of genetically modified food causes that countries in the world government greatly pays close attention to, and therefore launches respectively the tag control system and the entry and exit product reporting system of genetically modified food.The requirement of United Nations and European Union is more strict, the regulation genetically modified food all needs to detect monitoring from studying, produce, transport, store, be sold to all links such as import and export, therefore an important process that becomes each main trading country check is identified in the detection of transgenic product, and the relevant stdn detection architecture of foundation, (referring to Zhu Guangfu, countries in the world are to the attitude and the management of genetically modified food, oil and foodstuffs science and technology to be adapted to various methods for detecting transgenic foods, 2001,3:1-5).
Use the transgene component in the PCR method detection deep-processed food, the ultimate challenge that is faced is the template DNA composition that how to extract from numerous highly processed goods effectively as the PCR reaction, the difficulty of its DNA extraction is that not only DNA is seriously damaged in the course of processing, and the factors such as the physics that produces in the course of processing, chemistry or biology also can have influence on the DNA quality.All kinds of different finely processed product that different work programs produces, the residual volume of DNA differs widely in the palliating degradation degree of its DNA and the product, therefore exploitation is the key that the deep-processed food transgene component detects at the template DNA extractive technique that the variant production transgenosis detects.
Summary of the invention
Present method is intended to set up from the soy sauce that processes with genetically engineered soybean extracts template DNA, and the PCR that is used for transgene component detects.
The objective of the invention is to overcome deficiency of the prior art, a kind of method of extracting the DNA that is used for the transgene component detection from soy sauce is provided, it can be quick, easy, reliably, expeditiously from being the class deep processing fermented products such as soy sauce that process of raw material template DNA to be extracted with the soybean, the qualitative detection that is used for transgene component is with the reliability of guaranteeing to detect.
Present method, must be removed the caramel colorant molecule before formally carrying out DNA extraction effectively at the high characteristics of caramel colorant content in the soy sauce.For this reason in adding the The pretreatment process, utilize the liquid soy sauce of dehydrated alcohol low-temperature sludge, effective constituent is wherein all precipitated, use 0.1mol/L Tris (pH8.0) solution dissolution precipitation again, on magnetic stirring apparatus, stir, but the thorough washing precipitation is to remove small-molecule substances such as caramel colorant and salt, and for the subsequent DNA leaching process reduces the heavy burdens, this is that DNA extraction is necessary.
Technical scheme of the present invention is summarized as follows:
Extract the method for the DNA that is used for the transgene component detection from soy sauce, it is made up of following step:
1. get soy sample 30mL, dehydrated alcohol with 1-3 times of soy sample volume, be placed on-20 ℃ of precipitations 5-15 minute, precipitation firmly shakes up with the 0.1mol/L Tris-HCl solution dissolving of the pH=8.0 of 0.5-1.5 times of soy sample volume, all be transferred in the 100mL beaker, on magnetic stirring apparatus, stirred 2 hours 12000 rev/mins of room temperatures, centrifugal 10 minutes, abandon supernatant, precipitation is used for DNA extraction.
2. whole precipitations that step 1 is obtained add 10-15mL and extract damping fluid through the bromohexadecane base Trimethylamine 99 of 65 ℃ of preheatings, mixing, insulation is 1-1.5 hour in 65 ℃ of water-baths, during put upside down mixing gently once every 10 minutes.
3. taking-up is left standstill and is cooled to 20-25 ℃, 12000 rev/mins, centrifugal 12 minutes, gets supernatant.
4. adding and the isopyknic volume ratio of the resulting supernatant liquor of step 3 are 24: 1 trichloromethane: primary isoamyl alcohol, and slowly put upside down and shake up 5 minutes, 10000 rev/mins, centrifugal 10 minutes, get supernatant, add 24: 1 trichloromethane of equal-volume again: the primary isoamyl alcohol extracting is once got supernatant.
5. add the bromohexadecane base Trimethylamine 99 precipitated liquid of 2 times of volumes, shake up, 20-25 ℃ left standstill 50-90 minute, and 12000 rev/mins, centrifugal 10 minutes, abandon supernatant, stay precipitation.
6. precipitation adds 1.2mol/L NaCl solution 1mL, dissolution precipitation, and 20-25 ℃ left standstill 5 minutes, added 24: 1 trichloromethane of equal-volume: primary isoamyl alcohol, put upside down and shake up 1 minute, 12000 rev/mins, centrifugal 10 minutes, get supernatant.
7. the 3mol/L sodium acetate and the isopyknic 4 ℃ of pre-cold isopropanols that add the institute's supernatant liquor of getting 1/10 volume, slow mixing, 4 ℃ of standing over night, 14000 rev/mins, centrifugal 15 minutes, abandon supernatant, stay precipitation.
8. with the 70% washing with alcohol precipitation of 400 μ L of 4 ℃ of precoolings, remove residual ethanol, drying precipitated, obtain the DNA precipitation.
9. add the deionized water of 25-35 μ L sterilization, the DNA precipitation that dissolving is extracted from soy sauce.
The invention has the beneficial effects as follows: effectively the caramel colorant in the soy sauce is removed, for the subsequent DNA extraction eases off the pressure, satisfied the requirement that transgene component detects, this method is quick, easy, high specificity, the efficient height, being suitable for the soybean is that the PCR of transgene component in the fermented products such as soy sauce that process of raw material detects.
Description of drawings
Endogenous special pcr amplification result (the 1.DNA Marker of Fig. 1 soy sauce DNA with reference to the Lectin gene; 2. soybean seeds; 3. soy sauce)
Embodiment
1. material and reagent
1) material: soy sample is available from the supermarket, contrasts to be soybean seeds.
2) extract reagent:
DNA extraction damping fluid (1L): dissolving bromohexadecane base Trimethylamine 99 20g in the 1000ml water, Tris 12.114g, NaCl 81.816g, Na
2EDTA 7.444g, the PVP[polyvinylpyrrolidone] 20g, autoclaving.
Bromohexadecane base Trimethylamine 99 precipitated liquid (1L): dissolving bromohexadecane base Trimethylamine 99 5g in the 1000ml water, NaCl2.34g, autoclaving.
Sodium chloride solution (1.2mol/L): dissolving NaCl 70g in the 1000ml water, autoclaving.
Sodium acetate solution (3mol/L): dissolving 408.1g sodium acetate trihydrate in the 800ml water, transfer pH to 5.2 with glacial acetic acid, add water and be settled to 1L, autoclaving.
Tris-HCl solution (0.1mol/L, pH8.0): with 800ml water dissolution 12.114g Tris, be cooled to room temperature, transfer pH to 8.0, be settled to 1L with concentrated hydrochloric acid.
TE damping fluid: Tris 0.01mol/L, Na
2EDTA 1.0mmol/L transfers pH to 8.0, autoclaving with hydrochloric acid.
Trichloromethane: primary isoamyl alcohol (24: 1); Dehydrated alcohol; 70% ethanol; Virahol; The deionization sterilized water.
2.DNA the flow process of extracting:
1) sample pretreatment
Get liquid state fermentation sample 30mL such as soy sauce, add 60mL dehydrated alcohol mixing, placed 10 minutes for-20 ℃, 4 ℃ then, 10000 rev/mins, centrifugal 10 minutes, abandon supernatant, in precipitation, add 30mL 0.1M Tris-HCl solution, firmly shake up, all be transferred in the 100mL beaker, on magnetic stirring apparatus, stirred 2 hours 12000 rev/mins of room temperatures, centrifugal 10 minutes, abandon supernatant, precipitation is used for DNA extraction.
2) learn from else's experience pretreated sample adds 10mL and extracts damping fluid, mixing through the bromohexadecane base Trimethylamine 99 of 65 ℃ of preheatings.65 ℃ of water-baths 1 hour, during put upside down mixing gently once every 10 minutes.
3) leave standstill and be cooled to room temperature, 12000 rev/mins, centrifugal 12 minutes, get supernatant.
4) add isopyknic trichloromethane+primary isoamyl alcohol and slowly put upside down and shake up 5 minutes, in 10000 rev/mins, centrifugal 10 minutes, get supernatant, add isopyknic trichloromethane+primary isoamyl alcohol and repeat extracting once, get supernatant.
5) the bromohexadecane base Trimethylamine 99 precipitated liquid of 2 times of volumes of adding shakes up, and room temperature left standstill 1 hour, and 12000 rev/mins, centrifugal 10 minutes, abandon supernatant, stay precipitation.
6) precipitation adds 1.2mol/L NaCl solution 1mL dissolution precipitation, and room temperature left standstill 5 minutes, and adding equal-volume trichloromethane+primary isoamyl alcohol is put upside down and shaken up 1 minute, in 12000 rev/mins, centrifugal 10 minutes, gets supernatant.
7) 3mol/L sodium acetate and isopyknic cold isopropanol of adding 1/10 volume, slow mixing, 4 ℃ of standing over night, 14000 rev/mins, centrifugal 15 minutes, abandon supernatant, stay precipitation.
8) the cold washing with alcohol precipitation of adding 400 μ L 70% is removed residual ethanol, and is drying precipitated.
9) add the DNA precipitation that the deionized water dissolving of 30 μ L sterilization extracts from soy sauce, get the template of 15 μ L as a PCR reaction; DNA extraction thing with the soybean seeds that compares dissolves with 100 μ LTE, and it is standby to put-20 ℃ of preservations.
10) whether extract the DNA that can be used for the PCR detection, adopt the qualitative TRAP of special endogenous reference gene PCR to identify.Experimental technique and condition are as follows:
1) the PCR primer sequence of special endogenous reference gene (seeing Table 1)
The PCR primer sequence of the special endogenous reference gene of table 1
Detect gene primer sequence amplification fragment length raw material crop
Just: 5 '-gcc ctc tac tcc acc ccc atc c-3 '
Lectin 118bp soybean
Instead: 5 '-gcc cat ctg caa gcc ttt ttg tg-3 '
2) the PCR reaction system of special endogenous reference gene is formed (seeing Table 2)
The amount of each reagent is suitably adjusted according to the cumulative volume of reaction system in the reaction system, and each reaction system should be provided with two parallel pipes.The reaction system that soybean oil DNA quality examination is adopted is identical.
Table 2 PCR detects reference reaction system (25 μ L system)
Reagent name adds the amount of PCR reaction system
10 * PCR reaction buffer (contains Mg
2+) 2.5 μ L
DNTP solution (every kind is 10mmol/L) 0.5 μ L
Forward primer (10pmol/ μ L) 2.5 μ L
Reverse primer (10pmol/ μ L) 2.5 μ L
Taq enzyme (2U/ μ L) 2.0 μ L
*Template DNA 10～15 μ L
Water is supplied reaction system, and making cumulative volume is 25 μ L
*: the dna profiling consumption of positive control sample is 1 μ L in the table.
3) the PCR reaction conditions of special endogenous reference gene (seeing Table 3)
The PCR of the special endogenous reference gene of table 3 detects the reference reaction condition
The gene sex change amplification cycles number of times of amplification extends at last
94℃，30s
62℃，30s
Lectin 96℃，2min 40 72℃，3min
72℃，25s
4) deposition condition
Sepharose (gel thawing back room temperature adds ethidium bromide when being placed into 60 ℃ of left and right sides, and content is 0.5g/mL) with 1 * TAE electrophoretic buffer preparation 2%.With PCR product and the sample-loading buffer uniform mixing of 10 μ L, join then in the gel pore in proportion, (3V/cm～5V/cm) carry out electrophoresis, electrophoresis time is 40min～60min to select suitable voltage.Place sepharose on the gel imaging instrument or observation and photograph on the ultraviolet transilluminator.
Below in conjunction with specific embodiment the present invention is further described.
Embodiment 1:
The naughty big soy sauce of washing in a pan the production of larger food company limited with Chinese-foreign joint Shanghai is a material, adopts the DNA in present method extraction soy sauce,
1) gets liquid state fermentation sample 30mL such as soy sauce, add 60mL dehydrated alcohol mixing, placed 10 minutes for-20 ℃, 4 ℃ then, 10000 rev/mins, centrifugal 10 minutes, abandon supernatant, in precipitation, add 30mL 0.1M Tris-HCl solution, firmly shake up, all be transferred in the 100mL beaker, on magnetic stirring apparatus, stirred 2 hours room temperature, 12000 rev/mins, centrifugal 10 minutes, abandon supernatant, precipitation is used for DNA extraction.
2) learn from else's experience pretreated sample adds 10mL and extracts damping fluid through the bromohexadecane base Trimethylamine 99 of 65 ℃ of preheatings, mixing, 65 ℃ of water bath heat preservations 1 hour, during put upside down mixing gently once every 10 minutes.
3) leave standstill and be cooled to room temperature, 12000 rev/mins, centrifugal 12 minutes, get supernatant.
4) add isopyknic trichloromethane+primary isoamyl alcohol and slowly put upside down and shake up 5 minutes, 10000 rev/mins, centrifugal 10 minutes, get supernatant, add isopyknic trichloromethane+primary isoamyl alcohol and repeat extracting once, get supernatant.
5) the bromohexadecane base Trimethylamine 99 precipitated liquid of 2 times of volumes of adding shakes up, and room temperature left standstill 1 hour, and 12000 rev/mins, centrifugal 10 minutes, abandon supernatant, stay precipitation.
6) precipitation adds 1.2mol/LNaCl solution 1mL dissolution precipitation, and room temperature left standstill 5 minutes, and adding equal-volume trichloromethane+primary isoamyl alcohol is put upside down and shaken up 1 minute, 12000 rev/mins, centrifugal 10 minutes, gets supernatant.
7) 3mol/L sodium acetate and isopyknic cold isopropanol of adding 1/10 volume, slow mixing, 4 ℃ of standing over night, 14000 rev/mins, centrifugal 15 minutes, abandon supernatant, stay precipitation.
8) the cold washing with alcohol precipitation of adding 400 μ L 70% is removed residual ethanol, and is drying precipitated.
9) add the DNA precipitation of extracting in the deionized water dissolving soy sauce of 30 μ L sterilization, get the template of 15 μ L as a PCR reaction; DNA extraction thing with the soybean seeds that compares dissolves with 100 μ L TE, and it is standby to put-20 ℃ of preservations.
Embodiment 2
1) gets liquid state fermentation sample 30mL such as soy sauce, add 30mL dehydrated alcohol mixing, placed 5 minutes for-20 ℃, 4 ℃ then, 10000 rev/mins, centrifugal 10 minutes, abandon supernatant, in precipitation, add 15mL 0.1M Tris-HCl solution, firmly shake up, all be transferred in the 100mL beaker, on magnetic stirring apparatus, stirred 2 hours 12000 rev/mins of room temperatures, centrifugal 10 minutes, abandon supernatant, precipitation is used for DNA extraction.
2)-4) step is with embodiment 1.
5) the bromohexadecane base Trimethylamine 99 precipitated liquid of 2 times of volumes of adding shakes up, and room temperature left standstill 50 minutes, and 12000 rev/mins, centrifugal 10 minutes, abandon supernatant, stay precipitation.
6)-8) step is with embodiment 1.
9) add the DNA precipitation of extracting in the deionized water dissolving soy sauce of 25 μ L sterilization, get the template of 15 μ L as the PCR reaction; DNA extraction thing with the soybean seeds that compares dissolves with 100 μ L TE, and it is standby to put-20 ℃ of preservations.
Embodiment 3
1) gets liquid state fermentation sample 30mL such as soy sauce, add 90mL dehydrated alcohol mixing, placed 15 minutes for-20 ℃, 4 ℃ then, 10000 rev/mins, centrifugal 10 minutes, abandon supernatant, in precipitation, add 45mL 0.1M Tris-HCl solution, firmly shake up, all be transferred in the 100mL beaker, on magnetic stirring apparatus, stirred 2 hours 12000 rev/mins of room temperatures, centrifugal 10 minutes, abandon supernatant, precipitation is used for DNA extraction.
2)-4) step is with embodiment 1.
5) the bromohexadecane base Trimethylamine 99 precipitated liquid of 2 times of volumes of adding shakes up, and room temperature left standstill 90 minutes, and 12000 rev/mins, centrifugal 10 minutes, abandon supernatant, stay precipitation.
6)-8) step is with embodiment 1.
9) add the DNA precipitation of extracting in the deionized water dissolving soy sauce of 35 μ L sterilization, get the template of 15 μ L as the PCR reaction; DNA extraction thing with the soybean seeds that compares dissolves with 100 μ L TE, and it is standby to put-20 ℃ of preservations.
Embodiment 4
1) gets liquid state fermentation sample 30mL such as soy sauce, add 50mL dehydrated alcohol mixing, place after 12 minutes for-20 ℃, 4 ℃ then, 10000 rev/mins, centrifugal 10 minutes, abandon supernatant, in precipitation, add 35mL 0.1M Tris-HCl solution, firmly shake up, all be transferred in the 100mL beaker, on magnetic stirring apparatus, stirred 2 hours 12000 rev/mins of room temperatures, centrifugal 10 minutes, abandon supernatant, precipitation is used for DNA extraction.
2)-4) step is with embodiment 1.
5) the bromohexadecane base Trimethylamine 99 precipitated liquid of 2 times of volumes of adding shakes up, and room temperature left standstill 70 minutes, and 12000 rev/mins, centrifugal 10 minutes, abandon supernatant, stay precipitation.
6)-8) step is with embodiment 1.
9) add the DNA precipitation of extracting in the deionized water dissolving soy sauce of 25 μ L sterilization, get the template of 15 μ L as the PCR reaction; DNA extraction thing with the soybean seeds that compares dissolves with 100 μ L TE, and it is standby to put-20 ℃ of preservations.
Embodiment 5
1) gets liquid state fermentation sample 30mL such as soy sauce, add 30mL dehydrated alcohol mixing, placed 12 minutes for-20 ℃, 4 ℃ then, 10000 rev/mins, centrifugal 10 minutes, abandon supernatant, in precipitation, add 35mL 0.1M Tris-HCl solution, firmly shake up, all be transferred in the 100mL beaker, on magnetic stirring apparatus, stirred 2 hours 12000 rev/mins of room temperatures, centrifugal 10 minutes, abandon supernatant, precipitation is used for DNA extraction.
2)-4) step is with embodiment 1.
5) the bromohexadecane base Trimethylamine 99 precipitated liquid of 2 times of volumes of adding shakes up, and room temperature left standstill 70 minutes, and 12000 rev/mins, centrifugal 10 minutes, abandon supernatant, stay precipitation.
6)-8) step is with embodiment 1.
9) add the DNA precipitation of extracting in the deionized water dissolving soy sauce of 35 μ L sterilization, get the template of 15 μ L as the PCR reaction; DNA extraction thing with the soybean seeds that compares dissolves with 100 μ L TE, and it is standby to put-20 ℃ of preservations.
Claims (6)
1. extract the method for the DNA that is used for the transgene component detection from soy sauce, it is made up of following step:
(1) gets soy sample 30mL, dehydrated alcohol with 1-3 times of soy sample volume was placed on-20 ℃ of precipitations 5-15 minute, and precipitation is dissolved with the 0.1mol/L Tris-HCl solution of the pH=8.0 of 0.5-1.5 times of soy sample volume, firmly shake up, all be transferred in the 100mL beaker, on magnetic stirring apparatus, stirred 2 hours room temperature, 12000 rev/mins, centrifugal 10 minutes, abandon supernatant, precipitation is used for DNA extraction;
(2) whole precipitations that step (1) is obtained add 10-15mL and extract damping fluid through the bromohexadecane base Trimethylamine 99 of 65 ℃ of preheatings, mixing, insulation is 1-1.5 hour in 65 ℃ of water-baths, during put upside down mixing gently once every 10 minutes;
(3) taking-up is left standstill and is cooled to 20-25 ℃, 12000 rev/mins, centrifugal 12 minutes, gets supernatant;
(4) adding and the isopyknic volume ratio of the resulting supernatant liquor of step (3) they are 24: 1 trichloromethane: primary isoamyl alcohol, and slowly put upside down and shake up 5 minutes, 10000 rev/mins, centrifugal 10 minutes, get supernatant, add 24: 1 trichloromethane of equal-volume again: the primary isoamyl alcohol extracting is once got supernatant;
(5) the bromohexadecane base Trimethylamine 99 precipitated liquid of 2 times of volumes of adding shakes up, and 20-25 ℃ left standstill 50-90 minute, and 12000 rev/mins, centrifugal 10 minutes, abandon supernatant, stay precipitation;
(6) precipitation adds 1.2mol/LNaCl solution 1mL, dissolution precipitation, and 20-25 ℃ left standstill 5 minutes, added 24: 1 trichloromethane of equal-volume: primary isoamyl alcohol, put upside down and shake up 1 minute, 12000 rev/mins, centrifugal 10 minutes, get supernatant;
(7) add 3mol/L sodium acetate and isopyknic 4 ℃ of pre-cold isopropanols of the institute's supernatant liquor of getting 1/10 volume, slow mixing, 4 ℃ of standing over night, 14000 rev/mins, centrifugal 15 minutes, abandon supernatant, stay precipitation;
(8) with the 70% washing with alcohol precipitation of 400 μ L of 4 ℃ of precoolings, remove residual ethanol, drying precipitated, obtain the DNA precipitation;
(9) add the deionized water that 25-35 μ L sterilizes, the DNA precipitation that dissolving is extracted from soy sauce.
2. the method for extracting the DNA that is used for the transgene component detection from soy sauce according to claim 1, it is characterized in that: dehydrated alcohol is 2 times of soy sauce volumes in the described step (1).
3. the method for extracting the DNA that is used for the transgene component detection from soy sauce according to claim 1, it is characterized in that: be 10 minutes-20 ℃ of storage periods in the described step (1).
4. the method be used for the DNA that transgene component detects of extracting from soy sauce according to claim 1 is characterized in that: the solution of 0.1mol/L Tris-HCl that described step (1) is used for the pH=8.0 of dissolution precipitation is 1 times of soy sauce volume.
5. the method for extracting the DNA that is used for the transgene component detection from soy sauce according to claim 1, it is characterized in that: the time of repose in the described step (5) is 60 minutes.
6. the method for extracting the DNA that is used for the transgene component detection from soy sauce according to claim 1 is characterized in that: the volume of sterilization deionized water is 30 μ L in the described step (9).
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101100479B (en) * | 2007-08-03 | 2011-03-30 | 安徽大学 | Method for extracting residual DNA in edible vegetable oil |
| CN102242114A (en) * | 2011-02-23 | 2011-11-16 | 中国检验检疫科学研究院 | Method for extracting total DNA from soy and application thereof |
| CN101659998B (en) * | 2009-09-23 | 2012-01-11 | 中山大学 | Kit for detecting soybean transgene components in soy and use method thereof |
| CN107365767A (en) * | 2017-09-15 | 2017-11-21 | 广东美格基因科技有限公司 | A kind of method that DNA is extracted from soy sauce |
-
2004
- 2004-07-28 CN CN 200410020177 patent/CN1597983A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101100479B (en) * | 2007-08-03 | 2011-03-30 | 安徽大学 | Method for extracting residual DNA in edible vegetable oil |
| CN101659998B (en) * | 2009-09-23 | 2012-01-11 | 中山大学 | Kit for detecting soybean transgene components in soy and use method thereof |
| CN102242114A (en) * | 2011-02-23 | 2011-11-16 | 中国检验检疫科学研究院 | Method for extracting total DNA from soy and application thereof |
| CN102242114B (en) * | 2011-02-23 | 2013-05-08 | 中国检验检疫科学研究院 | Method for extracting total DNA from soy and application thereof |
| CN107365767A (en) * | 2017-09-15 | 2017-11-21 | 广东美格基因科技有限公司 | A kind of method that DNA is extracted from soy sauce |
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