SN/T 2051-2008 PDF English
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Determination of bovine, ovine, porcine-derived materials in food, cosmetic and feed-Real-time PCR method
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SN/T 2051-2008: Determination of bovine, ovine, porcine-derived materials in food, cosmetic and feed-Real-time 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/SNT2051-2008
SN
Entry-Exit Inspection and Quarantine Standards
of the People’s Republic of China
SN/T 2051-2008
Determination of bovine, ovine, porcine-derived materials in
food, cosmetic and feed - Real-time PCR method
Issued on. APRIL 29, 2008
Implemented on. NOVEMBER 01, 2008
Issued by. General Administration of Quality Supervision, Inspection and
Quarantine of the People's Republic of China
SN/T 2051-2008
Table of Contents
Foreword... 3
1 Scope... 4
2 Normative references... 4
3 Terms, definitions and abbreviations... 4
4 Principle... 5
5 Materials and reagents... 6
6 Sampling... 7
7 Operation methods... 7
8 Judgment criteria... 8
9 Low-limit of determination... 10
Annex A... 11
Annex B... 15
Annex C... 16
Annex D... 17
Annex E... 18
Annex F... 19
SN/T 2051-2008
Foreword
Annex F is normative; Annexes A, B, C, D and E are informative.
This Standard shall be under the jurisdiction of Certification and Accreditation Administration of the
People’s Republic of China.
Drafting organizations of this standard. Liaoning Entry-Exit Inspection and Quarantine Bureau,
Takara Biotechnology (Dalian) Co., Ltd. AND Chinese Academy of Inspection and Quarantine.
Main drafters of this standard. Cao Jijuan, Li Jingquan, Zhen Qiuyue, Yu Aili, Chen Yin, Yao Shan
and Xie Yan.
This is the first-time to publish this entry-exit inspection and quarantine standard.
SN/T 2051-2008
Determination of bovine, ovine, porcine-derived materials in food, cosmetic and
feed - Real-time PCR method
1 Scope
This standard specifies the real-time PCR detection methods for bovine, ovine (including sheep
and goat), porcine-derived materials in food, cosmetic and feed.
This standard is applicable to the identification detection of bovine, ovine (including sheep and
goat), porcine-derived materials in food, cosmetic and feed.
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 subsequent modifications (including all
corrections) or revisions made thereafter do not apply to this standard. However, the parties who
reach an agreement according to this standard are encouraged to study whether the latest
versions of these documents are applicable. For the undated documents so quoted, the latest
versions (including all modification sheets) apply to this document.
GB/T 6682 Water for analytical laboratory use - Specification and test methods (GB/T
6682-1992, neq ISO 3696.1987)
GB 19489 Laboratories - General requirements for biosafety
3 Terms, definitions and abbreviations
For the purpose of this standard, the following terms, definitions and abbreviations apply.
3.1 Terms and definitions
3.2 Abbreviations
3.2.1
DNA deoxyribonuleic acid
3.2.2
PCR polymerase chain reaction.
4 Principle
This standard adopts TaqMan real-time PCR technology; it identifies the variety of animal derived
materials based on the discrepancy of polymorphism in mitochondrial DNA (COX I) of animal
species. The multi-color fluorescence detection and multi-PCR method are applied in this
standard.
5 Materials and reagents
5.1 Instrumentation and materials
5.1.4 Vibrator.
5.1.5 Refrigerator (2°C ~ 8°C and -20°C or -80°C).
5.1.6 Micro-scale adjustable pipette and matched sucker (10 μL, 100 μL, 1 000 μL).
5.1.7 Real-time PCR reaction pipe.
5.1.8 Centrifuge pipe.
5.1.9 Water bath.
5.1.10 Heater.
5.1.11 Mobile ultraviolet lamp.
5.1.12 Magnetic shelf.
5.2 Reagents
Unless otherwise specified, all the reagents are analytically pure. The water is grade-1 water as
prescribed in GB/T 6682.All the reagents shall be stored in the container which will not pollute
DNA enzyme.
5.2.1 The composition, functions and use attentions of genomic DNA extraction kit 1) are
prescribed in Annex A.
6 Sampling
6.1 Sampling tools
The sampling tools such as scissors, tweezers and prod shall be sterilized at 180°C±2°C for 2h.
6.2 Sample collection
The sample to be tested shall be packed in the disposable plastic bag or other sterilized
containers, be given a number, and then delivered to the laboratory.
7 Operation methods
7.1 Laboratory instrumentation and management
The instrumentation in the laboratory and their management are prescribed in Annex F.
7.2 Preparation of template DNA sample
7.2.1 The template DNA sample shall be prepared at the sample preparation area. The
composition, functions and use attentions are prescribed in Annex A.
7.3 Detection
7.3.1 Preparation of amplification reagents
The preparation of amplification reagent shall be carried out in the reaction mixture preparation
area. To ensure the accuracy of detection results, when undergoing the detection by the real
sample, blank control, negative control, and positive control experiment shall be set up.
7.3.2 Sample addition
The addition of sample shall be carried out in the sampling area. Add the prepared template DNA
solution to each setup PCR reaction pipe. Enclose the pipe and centrifuge it for 5s ~10s.
8 Judgment criteria
8.1 Setup of result analysis conditions
The detection results can be directly read. The principle for the setup of valve values shall be
adjusted based on the noise conditions of instrumentation.
8.2 Quality control criteria
8.2.1 In the case of negative control experiment for bovine, ovine, porcine-derived materials, if
HEX fluorescence signal is detected and the typical amplification curve appears, Ct value shall be
less than 28.0 but no FAM fluorescence signal is detected.
8.3 Judgment criteria and specifications
8.3.1 Valid principles
Ct value less than or equal to 35 is regarded as valid value and Ct value greater than 35 is
regarded as invalid value.
8.3.3 Result description
The result is described as “bovine, ovine, porcine-derived materials detected” or “bovine, ovine,
porcine-derived materials not detected”.
9 Low-limit of determination
Under the conditions as described above, the low-limit of the determination by this method is
0.1%.
Annex A
(Informative)
Composition, Extraction Methods and Precautions for Genomic DNA Extraction
Kit from Food, Cosmetic and Feed
A.1 Genomic DNA extraction kit in solid food and feed
Take the general-use genomic DNA extraction kit produced by Takara Bio (Dalian) Co., Ltd. (Art.
No. DV811) for example.
A.1.1 Composition of kit
The kit consists of reagent and pillar. Each kit can handle the treatment of 50 samples which
consists of the following constituents.
A.1.5 Use attentions
A.1.5.1 Cross contamination shall be avoided when handling the sample.
A.1.5.2 In the case of using the ground sample with the addition of liquid nitrogen, the liquid
nitrogen shall be added to ensure the extracted genomic DNA will not be dissolved.
A.1.5.3 In the case of two-phase separation, when water phase solution (lowest layer colorless) is
transferred to the filter bowl, please discard the colored organic phase on the top layer or
otherwise it will restrain DNA binding to DNA preparation membrane. Inter-phase sedimentation
shall not be considered because it will be discarded during filtration.
A.1.5.4 If the partial reagent contains irritant compounds, please wear rubber gloves and glasses
during operation, so as to prevent the chemicals contacting skin and eyes. The operation shall be
carried out in the well-ventilated chemical hood. In the event that the chemicals contact the skin or
eyes, wash the skin or eyes with clean water and immediately send the injury to hospital for
medical care if necessary.
A.1.5.5 In the case of long-term storage of genomic DNA, it is suggested to be stored in the
eluent.
SN/T 2051-2008
SN
Entry-Exit Inspection and Quarantine Standards
of the People’s Republic of China
SN/T 2051-2008
Determination of bovine, ovine, porcine-derived materials in
food, cosmetic and feed - Real-time PCR method
Issued on. APRIL 29, 2008
Implemented on. NOVEMBER 01, 2008
Issued by. General Administration of Quality Supervision, Inspection and
Quarantine of the People's Republic of China
SN/T 2051-2008
Table of Contents
Foreword... 3
1 Scope... 4
2 Normative references... 4
3 Terms, definitions and abbreviations... 4
4 Principle... 5
5 Materials and reagents... 6
6 Sampling... 7
7 Operation methods... 7
8 Judgment criteria... 8
9 Low-limit of determination... 10
Annex A... 11
Annex B... 15
Annex C... 16
Annex D... 17
Annex E... 18
Annex F... 19
SN/T 2051-2008
Foreword
Annex F is normative; Annexes A, B, C, D and E are informative.
This Standard shall be under the jurisdiction of Certification and Accreditation Administration of the
People’s Republic of China.
Drafting organizations of this standard. Liaoning Entry-Exit Inspection and Quarantine Bureau,
Takara Biotechnology (Dalian) Co., Ltd. AND Chinese Academy of Inspection and Quarantine.
Main drafters of this standard. Cao Jijuan, Li Jingquan, Zhen Qiuyue, Yu Aili, Chen Yin, Yao Shan
and Xie Yan.
This is the first-time to publish this entry-exit inspection and quarantine standard.
SN/T 2051-2008
Determination of bovine, ovine, porcine-derived materials in food, cosmetic and
feed - Real-time PCR method
1 Scope
This standard specifies the real-time PCR detection methods for bovine, ovine (including sheep
and goat), porcine-derived materials in food, cosmetic and feed.
This standard is applicable to the identification detection of bovine, ovine (including sheep and
goat), porcine-derived materials in food, cosmetic and feed.
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 subsequent modifications (including all
corrections) or revisions made thereafter do not apply to this standard. However, the parties who
reach an agreement according to this standard are encouraged to study whether the latest
versions of these documents are applicable. For the undated documents so quoted, the latest
versions (including all modification sheets) apply to this document.
GB/T 6682 Water for analytical laboratory use - Specification and test methods (GB/T
6682-1992, neq ISO 3696.1987)
GB 19489 Laboratories - General requirements for biosafety
3 Terms, definitions and abbreviations
For the purpose of this standard, the following terms, definitions and abbreviations apply.
3.1 Terms and definitions
3.2 Abbreviations
3.2.1
DNA deoxyribonuleic acid
3.2.2
PCR polymerase chain reaction.
4 Principle
This standard adopts TaqMan real-time PCR technology; it identifies the variety of animal derived
materials based on the discrepancy of polymorphism in mitochondrial DNA (COX I) of animal
species. The multi-color fluorescence detection and multi-PCR method are applied in this
standard.
5 Materials and reagents
5.1 Instrumentation and materials
5.1.4 Vibrator.
5.1.5 Refrigerator (2°C ~ 8°C and -20°C or -80°C).
5.1.6 Micro-scale adjustable pipette and matched sucker (10 μL, 100 μL, 1 000 μL).
5.1.7 Real-time PCR reaction pipe.
5.1.8 Centrifuge pipe.
5.1.9 Water bath.
5.1.10 Heater.
5.1.11 Mobile ultraviolet lamp.
5.1.12 Magnetic shelf.
5.2 Reagents
Unless otherwise specified, all the reagents are analytically pure. The water is grade-1 water as
prescribed in GB/T 6682.All the reagents shall be stored in the container which will not pollute
DNA enzyme.
5.2.1 The composition, functions and use attentions of genomic DNA extraction kit 1) are
prescribed in Annex A.
6 Sampling
6.1 Sampling tools
The sampling tools such as scissors, tweezers and prod shall be sterilized at 180°C±2°C for 2h.
6.2 Sample collection
The sample to be tested shall be packed in the disposable plastic bag or other sterilized
containers, be given a number, and then delivered to the laboratory.
7 Operation methods
7.1 Laboratory instrumentation and management
The instrumentation in the laboratory and their management are prescribed in Annex F.
7.2 Preparation of template DNA sample
7.2.1 The template DNA sample shall be prepared at the sample preparation area. The
composition, functions and use attentions are prescribed in Annex A.
7.3 Detection
7.3.1 Preparation of amplification reagents
The preparation of amplification reagent shall be carried out in the reaction mixture preparation
area. To ensure the accuracy of detection results, when undergoing the detection by the real
sample, blank control, negative control, and positive control experiment shall be set up.
7.3.2 Sample addition
The addition of sample shall be carried out in the sampling area. Add the prepared template DNA
solution to each setup PCR reaction pipe. Enclose the pipe and centrifuge it for 5s ~10s.
8 Judgment criteria
8.1 Setup of result analysis conditions
The detection results can be directly read. The principle for the setup of valve values shall be
adjusted based on the noise conditions of instrumentation.
8.2 Quality control criteria
8.2.1 In the case of negative control experiment for bovine, ovine, porcine-derived materials, if
HEX fluorescence signal is detected and the typical amplification curve appears, Ct value shall be
less than 28.0 but no FAM fluorescence signal is detected.
8.3 Judgment criteria and specifications
8.3.1 Valid principles
Ct value less than or equal to 35 is regarded as valid value and Ct value greater than 35 is
regarded as invalid value.
8.3.3 Result description
The result is described as “bovine, ovine, porcine-derived materials detected” or “bovine, ovine,
porcine-derived materials not detected”.
9 Low-limit of determination
Under the conditions as described above, the low-limit of the determination by this method is
0.1%.
Annex A
(Informative)
Composition, Extraction Methods and Precautions for Genomic DNA Extraction
Kit from Food, Cosmetic and Feed
A.1 Genomic DNA extraction kit in solid food and feed
Take the general-use genomic DNA extraction kit produced by Takara Bio (Dalian) Co., Ltd. (Art.
No. DV811) for example.
A.1.1 Composition of kit
The kit consists of reagent and pillar. Each kit can handle the treatment of 50 samples which
consists of the following constituents.
A.1.5 Use attentions
A.1.5.1 Cross contamination shall be avoided when handling the sample.
A.1.5.2 In the case of using the ground sample with the addition of liquid nitrogen, the liquid
nitrogen shall be added to ensure the extracted genomic DNA will not be dissolved.
A.1.5.3 In the case of two-phase separation, when water phase solution (lowest layer colorless) is
transferred to the filter bowl, please discard the colored organic phase on the top layer or
otherwise it will restrain DNA binding to DNA preparation membrane. Inter-phase sedimentation
shall not be considered because it will be discarded during filtration.
A.1.5.4 If the partial reagent contains irritant compounds, please wear rubber gloves and glasses
during operation, so as to prevent the chemicals contacting skin and eyes. The operation shall be
carried out in the well-ventilated chemical hood. In the event that the chemicals contact the skin or
eyes, wash the skin or eyes with clean water and immediately send the injury to hospital for
medical care if necessary.
A.1.5.5 In the case of long-term storage of genomic DNA, it is suggested to be stored in the
eluent.
......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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