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NY/T 674-2003: Detection of genetically modified plant organisms and derived products. DNA extraction and purification
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Detection of genetically modified plant organisms and derived products. DNA extraction and purification
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NY/T 674-2003
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Basic data | Standard ID | NY/T 674-2003 (NY/T674-2003) | | Description (Translated English) | Detection of genetically modified plant organisms and derived products. DNA extraction and purification | | Sector / Industry | Agriculture Industry Standard (Recommended) | | Classification of Chinese Standard | B20 | | Classification of International Standard | 65.020.99 | | Word Count Estimation | 4,412 | | Date of Issue | 2003-04-01 | | Date of Implementation | 2003-05-15 | | Quoted Standard | NY/T 672; NY/T 673 | | Summary | This standard specifies: transgenic plants and their products in the DNA extraction and purification methods and technical requirements. This standard applies to: DNA-extraction and purification of the transgenic plants and their products. | NY/T 674-2003: Detection of genetically modified plant organisms and derived products. DNA extraction and purification
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Detection of genetically modified plant organisms and derived products.DNA extraction and purification
ICS 65.020.99
B20
People's Republic of China Agricultural Industry Standard
Transgenic plants and their products testing
DNA extraction and purification
Released on.2003-04-01
Implementation of.2003-05-15
Published by the Ministry of Agriculture
Foreword
This standard was proposed by the Department of Science and Technology Education of the Ministry of Agriculture.
This standard was drafted. Institute of Biotechnology, Chinese Academy of Agricultural Sciences, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Shanghai Agricultural Sciences
Institute, Ministry of Agriculture Science and Technology Development Center, China Agricultural University.
The main drafters of this standard. Jia Shirong, Jin Yujun, Zhang Dabing, Peng Yufa, Huang Kunlun, Li Ning, Wang Qihuai, Luo Yunbo.
This standard was first published.
Transgenic plants and their products testing
DNA extraction and purification
1 Scope
This standard specifies the methods and technical requirements for DNA extraction and purification in transgenic plants and their products.
This standard applies to the extraction and purification of DNA from transgenic plants and their products.
2 Normative references
The terms in the following documents become the terms of this standard by reference to this standard. All dated references, followed by all
Modifications (not including errata content) or revisions do not apply to this standard, however, parties to agreements based on this standard are encouraged to study
Is it possible to use the latest version of these files? For undated references, the latest edition applies to this standard.
General requirements for the detection of NY/T 672 transgenic plants and their products
NY/T 673 transgenic plants and their products testing sampling
3 Principle
DNA is separated from the different components of the sample by physical and chemical methods. Discard samples using different purification methods
Protein, fat, polysaccharides and other secondary metabolites, and chloroform, isoamyl alcohol, isopropanol, ethanol and
Purified DNA is obtained by sodium acetate or the like.
4 reagents and solutions
Unless otherwise stated, only analytically pure reagents were used in the analysis; the prepared solutions were used after autoclaving.
4.1 cetyltrimethylammonium bromide (CTAB).
4.2 Tris (hydroxymethyl) aminomethane, Tris].
4.3 ethylenediaminetetraacetic acid (disodiumsalt, dihydrate, EDTA-Na·
2H2O).
4.4 E-mercaptoethanol.
4.5 Chloroform.
4.6 Isoamyl alcohol (isoamylalcohol).
4.7 Isopropyl alcohol (isopropylalcohol).
4.8 Trichloromethane + isoamyl alcohol = 24 + 1 (volume ratio).
4.9 76% ethanol solution. Take 760mL of absolute ethanol, and dilute to 1L with water.
4.10 70% ethanol solution. Take 700mL of absolute ethanol, and bring up to 1L with water.
4.11 CTAB extraction buffer I. Prepare CTAB extraction buffer I per liter, should add 46.75g chlorine in 800mL deionized water
Sodium, shake the container to completely dissolve the solute, then add 50mL 1mol/LTris-HCl (pH 8.0) [dissolve in 800mL deionized water
Solve 121.1g Tris, cool to room temperature and adjust the pH of the solution to 8.0 with concentrated hydrochloric acid (about 42mL thick)
Hydrochloric acid), add water to 1L, autoclave after dispensing], 20mL0.5mol/LEDTA (pH 8.0) [Add in 800mL water
186.1 g of disodium edetate (EDTA-Na·2H2O), vigorously stirred on a magnetic stirrer, using sodium hydroxide (NaOH)
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