BSI PD CEN/TS 17303:2019

$142.49

Foodstuffs. DNA barcoding of fish and fish products using defined mitochondrial cytochrome b and cytochrome c oxidase I gene segments

Published By	Publication Date	Number of Pages
BSI	2019	28

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Categories: 67.120.30 - Fish and fishery products, BSI

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This document describes a procedure for the identification of single fish and fish fillets to the level of genus or species. The identification of fish species is carried out by PCR amplification of either a segment of the mitochondrial cytochrome b gene (cytb) or the cytochrome c oxidase I gene (cox1, syn COI) or both, followed by sequencing of the PCR products and subsequent sequence comparison with entries in databases. The methodology allows the identification of a large number of commercially important fish species. The decision whether the cytb or cox1 gene segment or both are used for fish identification depends on the declared fish species, the applicability of the PCR method for the fish species and the availability of comparative sequences in the public databases. This method has been successfully validated on raw fish fillets, however, laboratory experience is available that it can also be applied to processed, e.g. cold smoked, hot smoked, salted, frozen, cooked, fried, deep-fried samples. This document is usually unsuitable for the analysis of highly processed foods, e.g. tins of fish, with highly degraded DNA where the fragment lengths are not sufficient for amplification of the targets. Furthermore, it is not applicable for complex fish products containing mixtures of two or more fish species.

PDF Catalog

PDF Pages	PDF Title
2	undefined
5	European foreword Introduction
7	1 Scope 2 Normative references 3 Terms and definitions
9	4 Principle 5 Reagents and materials 5.1 General
10	5.2 PCR reagents
11	6 Apparatus 7 Procedure 7.1 Sample preparation 7.2 DNA extraction 7.3 PCR 7.3.1 General
12	7.3.2 PCR setup
13	7.3.3 Temperature-time program 7.3.4 PCR controls
14	8 Evaluation 8.1 Evaluation of PCR products 8.2 Evaluation of the PCR results 8.3 Sequencing of PCR products
15	8.4 Evaluation of sequence data 8.5 Comparison of the sequence with public databases 8.5.1 General 8.5.2 Sequence comparison of cytb and/or cox1 DNA sequences with GenBank
16	8.5.3 Sequence comparison of cox1 DNA sequences with BOLD
17	9 Interpretation of database query results 10 Validation status and performance criteria 10.1 Collaborative study for the identification of fish species based on cytb sequence analysis
18	10.2 Collaborative study for the identification of fish species based on cox1 sequence analysis
20	11 Test report
21	Annex A (informative)Practical laboratory experiences with the amplificability of cytb or cox1 segments from tested fish species
25	Bibliography