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GB/T 38132-2019: Quantitative determination of genetically modified plants by digital PCR method Delivery: 9 seconds. True-PDF full-copy in English & invoice will be downloaded + auto-delivered via email. See step-by-step procedure Status: Valid
Similar standardsGB/T 38132-2019: Quantitative determination of genetically modified plants by digital PCR method---This is an excerpt. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.), auto-downloaded/delivered in 9 seconds, can be purchased online: https://www.ChineseStandard.net/PDF.aspx/GBT38132-2019NATIONAL STANDARD OF THE PEOPLE’S REPUBLIC OF CHINA ICS 07.080 A 40 Quantitative determination of genetically modified plants by digital PCR method ISSUED ON: OCTOBER 18, 2019 IMPLEMENTED ON: OCTOBER 18, 2019 Issued by: State Administration for Market Regulation; Standardization Administration of the People’s Republic of China. Table of ContentsForeword ... 3 1 Scope ... 4 2 Normative references ... 4 3 Terms and definitions, abbreviations ... 4 4 Principle ... 5 5 Reagents and materials ... 6 6 Instruments and equipment ... 7 7 Operation steps ... 7 8 Results analysis and presentation ... 9 Appendix A (Informative) Genetically modified plants specific amplified sequences ... 11 Quantitative determination of genetically modified plants by digital PCR method1 ScopeThis Standard specifies the quantitative determination of genetically modified plants by digital PCR method. This Standard applies to the digital PCR method for quantitative determination of genetically modified maize MON810, MON89034, MIR162, genetically modified soybean GTS-40-3-2, genetically modified rice KMD event, genetically modified cotton GHB119, and genetically modified oilseed rape RT73 in seeds and physically-processed seeds. The quantitative detection limit of this method is 0.1% (mass fraction).2 Normative referencesThe following documents are indispensable for the application of this document. For dated references, only the dated version applies to this document. For undated references, the latest edition (including all amendments) applies to this document. GB/T 6682, Water for analytical laboratory use - Specification and test methods GB/T 19495.1, Detection of genetically modified organisms and derived products - General requirements and definitions GB/T 19495.3, Detection of genetically modified organisms and derived products - Nucleic acid extraction GB/T 19495.7, Detection of genetically modified organisms and derived products - Methods for sampling and sample preparation SN/T 4853, Quantitative detection of genetically modified rice - Digital PCR3 Terms and definitions, abbreviations3.1 Terms and definitions The following terms and definitions are applicable to this document. 3.1.1 Event specific sequence The neighbor-joining sequence that is recombined and produced after the exogenous DNA fragment is inserted into the recipient crop genome. 3.2 Abbreviations The following abbreviations are applicable to this document. Adh-1: Alcohol dehydrogenase 1 Adhc: Alcohol dehydrogenase C gene PCR: Polymerase chain reaction PEP: Phosphoenolpyruvate carboxylase gene PLD: Phospholipase D gene4 PrincipleThe technical principle of digital PCR is to segment the original PCR reaction system, and then to amplify and detect all small reaction systems. Through the limited segmentation of the reaction system, the entire reaction system can be more tolerant to the nucleic acid inhibitor, and can more stably, accurately, and rapidly perform a precise identification of the genetically modified organism of the trace. At present, digital PCR includes chip-type digital PCR and droplet-type digital PCR. The chip-type digital PCR uses a microfluidic chip to realize the segmentation of the original reaction system; this segmentation method has the advantages of good stability and uniformity; but the test cost is relatively high. The droplet-type digital PCR generates tiny water-in-oil system to realize the segmentation of the reaction system; this segmentation method has fast response speed and lower segmentation cost. In order to achieve quantitative determination of genetically modified plants by digital PCR, this Standard directly obtains the copy number content of exogenous genes (event specific sequence) and endogenous genes of genetically modified plants through digital PCR amplification reactions. The ratio (percentage) of the copy number of the exogenous gene to the endogenous gene in the sample DNA is the relative percentage content of the corresponding genetically modified plants in the sample. 7.5.2 Digital PCR reaction procedures The digital PCR reaction procedures are shown in Tables 4 and 5. 7.5.3 Setting of the control digital PCR reaction A positive control, a negative control and a blank control shall be set in the test. Use the genomic DNA of genetically modified plants that contains event specific sequence as the positive control; use the genomic DNA of non-genetically modified plants that contains the same endogenous genes as a negative control; use water as a blank control. In each control PCR reaction system, except for the template, the remaining components and PCR reaction conditions are the same as 7.5.1 and 7.5.2.8 Results analysis and presentation8.1 Quality control Set the threshold limit according to the end-point fluorescence value of the negative control in the digital PCR results; the threshold limit shall effectively distinguish the negative reaction microcells from the positive reaction microcells in the same sample. The negative control has endogenous gene amplification; neither the negative control nor the blank control has exogenous gene amplification. The relative standard deviation of the amplification parallel test result (percentage of genetically modified events) shall be less than or equal to 25%. ......Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al. Tips & Frequently Asked Questions:Question 1: How long will the true-PDF of English version of GB/T 38132-2019 be delivered?Answer: The full copy PDF of English version of GB/T 38132-2019 can be downloaded in 9 seconds, and it will also be emailed to you in 9 seconds (double mechanisms to ensure the delivery reliably), with PDF-invoice.Question 2: Can I share the purchased PDF of GB/T 38132-2019_English with my colleagues?Answer: Yes. The purchased PDF of GB/T 38132-2019_English will be deemed to be sold to your employer/organization who actually paid for it, including your colleagues and your employer's intranet.Question 3: Does the price include tax/VAT?Answer: Yes. 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