SN/T 1204-2003 (SN/T1204-2003, SNT 1204-2003, SNT1204-2003)
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Protocol of the real-time PCR method for detecting genetically modified plants and their derived products
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Protocol of the real-time PCR for detecting genetically modified plants and their derived products
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| Standard ID | SN/T 1204-2003 (SN/T1204-2003) | | Description (Translated English) | Protocol of the real-time PCR for detecting genetically modified plants and their derived products | | Sector / Industry | Commodity Inspection Standard (Recommended) | | Classification of Chinese Standard | B16 | | Word Count Estimation | 8,851 | | Date of Issue | 2003-03-17 | | Date of Implementation | 2003-09-01 | | Quoted Standard | GB/T 6682; SN/T 1193; SN/T 1194 | | Drafting Organization | General Administration of Quality Supervision, Inspection and Quarantine Laboratory Animal and Plant Quarantine | | Administrative Organization | National Certification and Accreditation Administration Committee | | Proposing organization | National Certification and Accreditation Administration Committee | | Issuing agency(ies) | General Administration of Quality Supervision, Inspection and Quarantine of the People Republic of China | | Summary | This standard specifies the genetically modified plants and their processed products in the real-time PCR qualitative test methods. This standard applies to corn, soybeans, rapeseed, potato, tomato, cotton, tobacco and processed products in genetically modified real-time PCR qualitative detection test or any other test method a confirmatory test result is positive. |
SN/T 1204-2003
SN
INDUSTRY STANDARD
OF THE PEOPLE’S 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’s Republic of China.
The responsible drafting organizations. General Office of Animal & 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. 0.5µL, 2µL, 10µL, 20µL, 100µL, 200µL, 1000µL.
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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