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SN/T 1204-2003 English PDF (SNT1204-2003)

SN/T 1204-2003 English PDF (SNT1204-2003)

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SN/T 1204-2003: Protocol of the real-time PCR for detecting genetically modified plants and their derived products

This Standard specifies the real-time fluorescence PCR qualitative testing method of the genetically modified ingredients in plants and its derived products. This Standard applies to the qualitative detection of real-time fluorescence PCR OR the validation detection for those positive results detected by other detection methods - maize, soybeans, rapeseeds, potatoes, tomatoes, cottons, tobaccos and their deprived products.
SN/T 1204-2003
SN
INDUSTRY STANDARD
OF THE PEOPLE REPUBLIC OF CHINA
Protocol of the real-time PCR for detecting genetically
modified plants and their derived products
ISSUED ON. MARCH 17, 2003
IMPLEMENTED ON. SEPTEMBER 1, 2003
Issued by. General Administration of Quality Supervision, Inspection
and Quarantine of the People's Republic of China
Table of Contents
Foreword ... 3
1 Scope ... 4
2 Normative references ... 4
3 Abbreviations ... 4
4 Anti-pollution measures ... 6
5 Sampling and preparation ... 6
6 Experimental methods ... 6
7 Quality control of real-time fluorescence PCR qualitative detection 11 8 Result judgment and presentation ... 11
Foreword
This Standard was proposed and shall be under the jurisdiction of Certification and Accreditation Administration of the PEOPLE Republic of China.
The responsible drafting organizations. General Office of Animal and Plant Quarantine of General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China, Guangdong Entry-Exit Inspection and Quarantine Bureau of P. R. China, Liaoning Entry-Exit Inspection and Quarantine Bureau of P. R. China, Shenzhen Entry-Exit Inspection and Quarantine Bureau of P. R. China, Shanghai Entry-Exit Inspection and Quarantine Bureau of P. R. China.
The chief drafting staffs of this Standard. Zhu Shuifang, Qin Wen, Cao Jijuan, Zhang Guiming, Pan Liangwen, Huang Wensheng, and Chen Hongyun.
This Standard is the inspection and quarantine industry standard which is issued for the first time.
Protocol of the Real-Time PCR for Detecting Genetically
Modified Plants and Their Derived Products
1 Scope
This Standard specifies the real-time fluorescence PCR qualitative testing method of the genetically modified ingredients in plants and its derived products.
This Standard applies to the qualitative detection of real-time fluorescence PCR OR the validation detection for those positive results detected by other detection methods - maize, soybeans, rapeseeds, potatoes, tomatoes, cottons, tobaccos and their deprived products. 2 Normative references
The articles contained in the following documents have become part of this Standard when they are quoted herein. For the dated documents so quoted, all the modifications (excluding corrections) or revisions made thereafter shall not be applicable to this Standard. For the undated documents so quoted, the latest editions shall be applicable to this Standard.
GB/T 6682 Water for analytical laboratory use - Specification and test methods SN/T 1193 General requirements for the laboratories for gene detection and identification
SN/T 1194 Methods of sampling and preparation of samples for detection of genetically modified components in plants and their derived products
3 Abbreviations
The following abbreviations are applicable to this Standard.
3.1 AmpErase UNG Enzyme. uracil N-glycosylase.
3.2 Ct Value. C; cycle, t. threshold, the cycle number that the fluorescence signal in each reaction tube goes through when it reaches the set threshold value.
6.2.5 Nucleic Acid Protein Analyzer.
6.2.6 High-Speed Refrigerated Centrifuge, Desk Miniature Centrifuge, Mini Personal Centrifuge.
6.2.7 Low-Temperature Refrigerator, Freezer.
6.2.8 Water Purifier, Double Distilled Water Maker.
6.2.9 Vortex Oscillators.
6.2.10 Microsyringes. 0.5??L, 2??L, 10??L, 20??L, 100??L, 200??L, 1000??L.
6.2.11 Photochemical PCR Reaction Tube.
6.3 Main reagents
Unless otherwise specified, all reagents shall adopt the analytical pure reagent or the biochemical reagent.
6.3.1 Water used in the experiment. It shall comply with the grade 1 water as specified in GB/T 6682.
6.3.2 PCR Buffer solution.
6.3.3 Magnesium chloride (MgCl2).
6.3.4 dNTPs (dATP, dUTP, dCTP, dGTP).
6.3.5 UNG enzyme (Uracil N-glycosylase).
6.3.6 Taq enzyme.
6.3.7 Primer and probe
See Table 1 for primers and probe sequences used for real-time PCR detection for genetically modified plants and their derived products.
real-time fluorescence PCR amplification shall be re-done. If the Ct value is still less than 40 after repetitive amplification, and the results of negative control, positive control and blank control are normal, then it is judged that XXX gene is detected from the sample. If the Ct value is more than 40 after repetitive amplification, and the results of negative control, positive control and blank are normal, then it is judged that XXX gene is not detected from the sample.
8.2 Results presentation
XXX gene is detected.
XXX gene is not detected.

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