SIST-TS CEN ISO/TS 21569-10:2026

Horizontal methods for molecular biomarker analysis - Methods of analysis for the detection of genetically modified organisms and derived products - Part 10: Construct- and event-specific detection methods for genetically modified salmon expressing CS-GHc2 growth hormone (ISO/TS 21569 10:2026)

This document specifies procedures for the detection of a DNA sequence of a construct used to (genetically) enhance the growth of fish commonly found in aquaculture. The genetically modified AquAdvantage Atlantic salmon (Salmo salar) carries the construct expressing CS-GHc2 growth hormone and can be detected based on a real-time polymerase chain reaction (PCR) targeting either the border between the growth hormone coding sequence (CS-GHc2) of Oncorhynchus tshawytscha (Chinook salmon) and the antifreeze terminator (T-AFP) of (Macro-) Zoarces americanus (ocean pout), i.e. with the construct-specific method, or the border between the Atlantic salmon genomic DNA and the antifreeze promoter (P-AFP) of ocean pout, i.e. with the event-specific method. These methods can be applied to identify the genetically modified (GM) fish or for screening purposes.
This document is applicable for the analysis of DNA extracted from foodstuffs. It can also be suitable for the analysis of DNA extracted from other products such as feedstuffs. The application of these methods requires the extraction of an adequate amount of amplifiable DNA from the relevant matrix.

Horizontale Verfahren für die molekulare Biomarker-Analyse - Verfahren zum Nachweis von gentechnisch modifizierten Organismen und ihren Produkten - Teil 10: Konstrukt- und ereignisspezifische Nachweisverfahren für gentechnisch veränderten Lachs, der das Wachstumshormon CS-GHc2 exprimiert (ISO/TS 21569 10:2026)

Dieses Dokument legt Verfahren für den Nachweis der DNA-Sequenz eines Konstrukts fest, das zur (genetischen) Steigerung des Wachstums von Fischen eingesetzt wird, die häufig in Aquakultur gehalten werden. Der gentechnisch modifizierte Atlantische Lachs AquAdvantage (Salmo salar) ist Träger dieses Konstrukts und kann anhand einer Real-time-Polymerase-Kettenreaktion (PCR) erkannt werden, die entweder gegen die Grenze zwischen der das Wachstumshormon codierenden Sequenz (CS-GHc2) von Oncorhynchus tshawytscha (Königslachs) und dem Terminator des Antifrost-Proteins (T-AFP) von (Macro )Zoarces americanus (Meeres-Dickkopf) oder die Grenze zwischen der genomischen DNA des Atlantischen Lachses und dem Promotor des Antifrost-Proteins (P-AFP) des Meeres-Dickkopfes gerichtet ist, d. h. mithilfe des transformationsereignis-spezifischen Verfahrens. Diese Verfahren können zur Erkennung von gentechnisch modifizierten Fischen oder zu Screening-Zwecken angewendet werden.
Dieses Dokument ist auf die Analyse von DNA anwendbar, die aus Lebensmitteln extrahiert wurde. Es kann jedoch auch zur Analyse von DNA geeignet sein, die aus anderen Produkten, wie z. B. Futtermitteln, extrahiert wurde. Voraussetzung für die Anwendung dieser Verfahren ist die Extraktion einer ausreichenden Menge amplifizierbarer DNA aus der betreffenden Matrix.

Méthodes horizontales pour l'analyse de biomarqueurs moléculaires - Méthodes d'analyse pour la détection des organismes génétiquement modifiés et des produits dérivés - Partie 10: Méthodes de détection spécifiques à la construction et à l'événement pour le saumon génétiquement modifié exprimant l'hormone de croissance CS-GHc2 (ISO/TS 21569 10:2026)

Horizontalne metode za analizo molekularnih biomarkerjev - Metode analize za odkrivanje gensko spremenjenih organizmov in iz njih izpeljanih proizvodov - 10. del: Metode za specifično odkrivanje konstrukta in dogodka za gensko spremenjenega lososa, ki izraža rastni hormon CS-GHc2 (ISO/TS 21569 10:2026)

Ta dokument določa postopke za odkrivanje DNA zaporedja konstrukta, ki se uporablja za (genetsko) izboljšanje rasti rib, ki se običajno nahajajo v akvakulturi. Genetsko spremenjeni AquAdvantage atlantski losos (Salmo salar) nosi konstrukt, ki izraža rastni hormon CS-GHc2, in ga je mogoče zaznati na podlagi realnočasovne verižne reakcije s polimerazo (PCR), ki cilja bodisi na mejo med kodirnim zaporedjem rastnega hormona (CS-GHc2) Oncorhynchus tshawytscha (čavica) in terminatorjem proti zmrzovanju (T-AFP) (Macro-) Zoarces americanus (oceanski pout), tj. s konstruktspecifično metodo, bodisi na mejo med genomskim DNA atlantskega lososa in promotorjem proti zmrzovanju (P-AFP) oceanskega pouta, tj. z dogodkovno specifično metodo. Te metode se lahko uporabijo za identifikacijo genetsko spremenjenih (GS) rib ali za namene presejanja.
Ta dokument je uporaben za analizo DNA, ekstrahiranega iz živil. Prav tako je lahko primeren za analizo DNA, ekstrahiranega iz drugih proizvodov, kot so krmila. Uporaba teh metod zahteva ekstrakcijo zadostne količine amplificirajočega DNA iz ustrezne matrice.

General Information

• Status: Published
• Public Enquiry End Date: 26-Apr-2026
• Publication Date: 06-Jul-2026
• ICS: 67.050 - General methods of tests and analysis for food products
• ICS: 67.120.30 - Fish and fishery products
• Technical Committee: KŽP - Agricultural food products
• Current Stage: 6060 - National Implementation/Publication (Adopted Project)
• Start Date: 11-Jun-2026
• Due Date: 16-Aug-2026
• Completion Date: 07-Jul-2026
• Ref Project: CEN ISO/TS 21569-10:2026 - Horizontal methods for molecular biomarker analysis - Methods of analysis for the detection of genetically modified organisms and derived products - Part 10: Construct- and event-specific detection methods for genetically modified salmon expressing CS-GHc2 growth hormone (ISO/TS 21569 10:2026)

Relations

• Replaces: SIST EN ISO 21569:2005 - Foodstuffs - Methods of analysis for the detection of genetically modified organisms and derived products - Qualitative nucleic acid based methods (ISO 21569:2005)
• Effective Date: 01-Jul-2026
• Replaces: SIST EN ISO 21569:2005/A1:2013 - Foodstuffs - Methods of analysis for the detection of genetically modified organisms and derived products - Qualitative nucleic acid based methods - Amendment 1 (ISO 21569:2005/Amd 1:2013)
• Effective Date: 01-Jul-2026
• Referred By: SIST-TS CEN/TS 16707:2014 - Foodstuffs - Methods of analysis for the detection of genetically modified organisms and derived products - Polymerase chain reaction (PCR) based screening strategies
• Effective Date: 10-Jun-2026
• Referred By: SIST-TS CEN/TS 15568:2007 - Foodstuffs - Methods of analysis for the detection of genetically modified organisms and derived products - Sampling strategies
• Effective Date: 10-Jun-2026

Overview

SIST-TS CEN ISO/TS 21569-10:2026 is an international technical specification developed by SIST for the detection of genetically modified organisms (GMOs) and their derived products in food and feed, focusing specifically on genetically modified salmon expressing the CS-GHc2 growth hormone. As part of the broader ISO 21569 series on horizontal methods for molecular biomarker analysis, this standard details both construct-specific and event-specific real-time PCR detection techniques targeted at the AquAdvantage Atlantic salmon (Salmo salar). These methods are essential for regulatory compliance, food authenticity verification, and traceability in the aquaculture and food industry.

Key Topics

• Scope and Objectives

  • Outlines robust procedures for detecting a unique DNA construct introduced in Atlantic salmon to enhance growth.
  • Specifies real-time PCR-based methods that target:
    • The border between the Chinook salmon CS-GHc2 gene and ocean pout antifreeze terminator (construct-specific approach).
    • The border between Atlantic salmon genomic DNA and the ocean pout antifreeze promoter (event-specific approach).
  • Provides guidelines for sample preparation, DNA extraction, PCR setup, and criteria for result validation.

• Performance and Validation

  • Describes sensitivity and specificity criteria, including detection limits and false-positive/negative rates.
  • Emphasizes method validation through interlaboratory trials and the use of standard control samples.
  • Offers strict protocols to ensure result accuracy and reproducibility across food matrices and laboratory settings.

• Applicability

  • Intended primarily for analyzing DNA extracted from foodstuffs, but also suitable for feedstuffs and other relevant products.
  • Requires extraction of amplifiable DNA for successful application of the detection methods.

• Technical Requirements

  • Recommends use of certified reagents, apparatus, and consumables suitable for molecular biology.
  • Details procedural controls, including amplification controls and PCR inhibition checks, supporting analytical reliability.

Applications

The standardized detection methods outlined in SIST-TS CEN ISO/TS 21569-10:2026 are critical in several areas:

• Regulatory Compliance

  • Supports regulatory authorities in enforcing labeling and traceability requirements for genetically modified food and feed products containing GM salmon.
  • Facilitates market surveillance and official food control activities.

• Food Authenticity and Safety

  • Enables food industry stakeholders and testing laboratories to verify the presence or absence of GM salmon in processed and raw food products.
  • Assures consumers regarding the authenticity and transparency of seafood supply chains.

• Aquaculture and Feed Industry

  • Provides the aquaculture sector with a means to monitor and control the distribution of genetically modified salmon.
  • Permits quality assurance testing in compound feeds where GMO content must be detected and quantified.

• Import/Export Controls

  • Assists customs and border inspection authorities in screening imported and exported fish products for unauthorized GM content.

Related Standards

Users of SIST-TS CEN ISO/TS 21569-10:2026 may also benefit from referencing these standards for comprehensive molecular biomarker and GMO analysis:

• ISO 21569: Foodstuffs - Methods for detection of GMOs and derived products - Qualitative nucleic acid-based methods
• ISO 21571:2005: Foodstuffs - Nucleic acid extraction methods for GMO detection
• ISO 24276: General requirements and definitions for methods of GMO analysis
• ISO 16577: Vocabulary for molecular biomarker analytical methods in agriculture and food

By following SIST-TS CEN ISO/TS 21569-10:2026, stakeholders ensure reliable, harmonized, and internationally recognized approaches for the detection of genetically modified salmon and related products throughout the food and aquaculture industries.

Standards Content (Sample)

SLOVENSKI STANDARD
01-september-2026
Nadomešča:
SIST EN ISO 21569:2005
SIST EN ISO 21569:2005/A1:2013
Horizontalne metode za analizo molekularnih biomarkerjev - Analitske metode za
določanje gensko spremenjenih organizmov in njihovih proizvodov - 10. del:
Metode določanja, ki so specifične za konstrukte in dogodke gensko
spremenjenega lososa, ki izraža rastni hormon CS-GHc2 (ISO/TS 21569 10:2026)
Horizontal methods for molecular biomarker analysis - Methods of analysis for the
detection of genetically modified organisms and derived products - Part 10: Construct-
and event-specific detection methods for genetically modified salmon expressing CS-
GHc2 growth hormone (ISO/TS 21569 10:2026)
Horizontale Verfahren für die molekulare Biomarker-Analyse - Verfahren zum Nachweis
von gentechnisch modifizierten Organismen und ihren Produkten - Teil 10: Konstrukt-
und ereignisspezifische Nachweisverfahren für gentechnisch veränderten Lachs, der das
Wachstumshormon CS-GHc2 exprimiert (ISO/TS 21569 10:2026)
Méthodes horizontales pour l'analyse de biomarqueurs moléculaires - Méthodes
d'analyse pour la détection des organismes génétiquement modifiés et des produits
dérivés - Partie 10: Méthodes de détection spécifiques à la construction et à l'événement
pour le saumon génétiquement modifié exprimant l'hormone de croissance CS-GHc2
(ISO/TS 21569 10:2026)
Ta slovenski standard je istoveten z: CEN ISO/TS 21569-10:2026
ICS:
67.120.30 Ribe in ribji proizvodi Fish and fishery products
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

CEN ISO/TS 21569-10
TECHNICAL SPECIFICATION
SPÉCIFICATION TECHNIQUE
June 2026
TECHNISCHE SPEZIFIKATION
ICS 67.120.30 Supersedes EN ISO 21569:2005, EN ISO
21569:2005/A1:2013
English Version
Horizontal methods for molecular biomarker analysis -
Methods of analysis for the detection of genetically
modified organisms and derived products - Part 10:
Construct- and event-specific detection methods for
genetically modified salmon expressing CS-GHc2 growth
hormone (ISO/TS 21569 10:2026)
Méthodes horizontales pour l'analyse de biomarqueurs Horizontale Verfahren für die molekulare Biomarker-
moléculaires - Méthodes d'analyse pour la détection Analyse - Verfahren zum Nachweis von gentechnisch
des organismes génétiquement modifiés et des modifizierten Organismen und ihren Produkten - Teil
produits dérivés - Partie 10: Méthodes de détection 10: Konstrukt- und ereignisspezifische
spécifiques à la construction et à l'événement pour le Nachweisverfahren für gentechnisch veränderten
saumon génétiquement modifié exprimant l'hormone Lachs, der das Wachstumshormon CS-GHc2 exprimiert
de croissance CS-GHc2 (ISO/TS 21569 10:2026) (ISO/TS 21569 10:2026)
This Technical Specification (CEN/TS) was approved by CEN on 28 May 2026 for provisional application.

The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to
submit their comments, particularly on the question whether the CEN/TS can be converted into a European Standard.

CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS
available promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in
parallel to the CEN/TS) until the final decision about the possible conversion of the CEN/TS into an EN is reached.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and
United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2026 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN ISO/TS 21569-10:2026 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
This document (CEN ISO/TS 21569-10:2026) has been prepared by Technical Committee ISO/TC 34
"Food products" in collaboration with Technical Committee CEN/TC 275 “Food analysis - Horizontal
methods” the secretariat of which is held by DIN.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
This document supersedes EN ISO 21569:2005, EN ISO 21569:2005/A1:2013.
Any feedback and questions on this document should be directed to the users’ national standards
body/national committee. A complete listing of these bodies can be found on the CEN website.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the
following countries are bound to announce this Technical Specification: Austria, Belgium, Bulgaria,
Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland,
Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Republic of
North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and the
United Kingdom.
Endorsement notice
The text of ISO/TS 21569-10:2026 has been approved by CEN as CEN ISO/TS 21569-10:2026 without
any modification.
Technical
Specification
ISO/TS 21569-10
First edition
Horizontal methods for molecular
2026-05
biomarker analysis — Methods
of analysis for the detection of
genetically modified organisms and
derived products —
Part 10:
Construct- and event-specific
detection methods for genetically
modified salmon expressing CS-
GHc2 growth hormone
Méthodes horizontales pour l'analyse de biomarqueurs
moléculaires — Méthodes d'analyse pour la détection des
organismes génétiquement modifiés et des produits dérivés —
Partie 10: Méthodes de détection spécifiques à la construction et
à l'événement pour le saumon génétiquement modifié exprimant
l'hormone de croissance CS-GHc2
Reference number
ISO/TS 21569-10:2026(en) © ISO 2026

ISO/TS 21569-10:2026(en)
© ISO 2026
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on
the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address below
or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: [phone]
Email: [e-mail]
Website: www.iso.org
Published in Switzerland
ii
ISO/TS 21569-10:2026(en)
Contents Page
Foreword .iv
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 2
5 Reagents and materials . 2
6 Apparatus . 3
7 Procedure . 3
7.1 Preparation of test samples .3
7.2 DNA extraction .3
7.3 PCR setup.3
7.4 Temperature-time programme.4
8 Accept/reject criteria . 4
8.1 General .4
8.2 Detection .5
9 Validation status and performance criteria . 5
9.1 Sensitivity .5
9.2 Specificity .5
9.3 Robustness of the two methods .7
9.4 Interlaboratory trial for the construct-specific detection method .7
9.4.1 General .7
9.4.2 PCR efficiency and limit of detection (LOD ) .8
95 %
9.4.3 False-positive and false-negative rates for DNA samples (blind samples) .9
9.5 Interlaboratory trial for the event-specific detection method .10
9.5.1 General .10
9.5.2 Validation results (blind samples) . .11
9.6 Summary evaluation .11
10 Test report .11
Bibliography .12

iii
ISO/TS 21569-10:2026(en)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
bodies (ISO member bodies). The work of preparing International Standards is normally carried out through
ISO technical committees. Each member body interested in a subject for which a technical committee
has been established has the right to be represented on that committee. International organizations,
governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely
with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
The procedures used to develop this document and those intended for its further maintenance are described
in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the different types
of ISO document should be noted. This document was drafted in accordance with the editorial rules of the
ISO/IEC Directives, Part 2 (see www.iso.org/directives).
ISO draws attention to the possibility that the implementation of this document may involve the use of (a)
patent(s). ISO takes no position concerning the evidence, validity or applicability of any claimed patent
rights in respect thereof. As of the date of publication of this document, ISO had not received notice of (a)
patent(s) which may be required to implement this document. However, implementers are cautioned that
this may not represent the latest information, which may be obtained from the patent database available at
www.iso.org/patents. ISO shall not be held responsible for identifying any or all such patent rights.
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and expressions
related to conformity assessment, as well as information about ISO’s adherence to the World Trade
Organization (WTO) principles in the Technical Barriers to Trade (TBT), see www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 16,
Horizontal methods for molecular biomarker analysis, in collaboration with the European Committee for
Standardization (CEN) Technical Committee CEN/TC 275, Food analysis - Horizontal methods, in accordance
with the Agreement on technical cooperation between ISO and CEN (Vienna Agreement).
A list of all parts in the ISO 21569 series can be found on the ISO website.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.

iv
Technical Specification ISO/TS 21569-10:2026(en)
Horizontal methods for molecular biomarker analysis —
Methods of analysis for the detection of genetically modified
organisms and derived products —
Part 10:
Construct- and event-specific detection methods for
genetically modified salmon expressing CS-GHc2 growth
hormone
1 Scope
This document specifies procedures for the detection of a DNA sequence of a construct used to (genetically)
enhance the growth of fish commonly found in aquaculture. The genetically modified AquAdvantage Atlantic
salmon (Salmo salar) carries the construct expressing CS-GHc2 growth hormone and can be detected based
on a real-time polymerase chain reaction (PCR) targeting either the border between the growth hormone
coding sequence (CS-GHc2) of Oncorhynchus tshawytscha (Chinook salmon) and the antifreeze terminator
(T-AFP) of (Macro-) Zoarces americanus (ocean pout), i.e. with the construct-specific method, or the border
between the Atlantic salmon genomic DNA and the antifreeze promoter (P-AFP) of ocean pout, i.e. with the
event-specific method. These methods can be applied to identify the genetically modified (GM) fish or for
screening purposes.
This document is applicable for the analysis of DNA extracted from foodstuffs. It can also be suitable for the
analysis of DNA extracted from other products such as feedstuffs. The application of these methods requires
the extraction of an adequate amount of amplifiable DNA from the relevant matrix.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content constitutes
requirements of this document. For dated references, only the edition cited applies. For undated references,
the latest edition of the referenced document (including any amendments) applies.
ISO 16577, Molecular biomarker analysis — Vocabulary for molecular biomarker analytical methods in
agriculture and food production
ISO 21569, Foodstuffs — Methods of analysis for the detection of genetically modified organisms and derived
products — Qualitative nucleic acid based methods
ISO 21571:2005, Foodstuffs — Methods of analysis for the detection of genetically modified organisms and
derived products — Nucleic acid extraction
ISO 24276, Foodstuffs — Methods of analysis for the detection of genetically modified organisms and derived
products — General requirements and definitions
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 16577 apply.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp

ISO/TS 21569-10:2026(en)
— IEC Electropedia: available at https:// www .electropedia .org/
4 Principle
DNA is extracted from the test sample by applying a suitable method with performance characteristics in
accordance with ISO 21571:2005.
The DNA analysis consists of the following two parts:
a) verification of the amount, quality and amplifiability of the extracted DNA (not part of this method);
b) analysis using the real-time PCR detection methods targeting the CS-GHc2 - T-AFP construct inserted in
the AquAdvantage salmon genome (construct-specific method, Accession AY594644, 75 bp amplicon)
[1][2]
or targeting the border between Atlantic salmon genome and P-AFP event in the AquAdvantage
[3][4]
salmon genome (event-specific method, 156 bp amplicon) .
Detection of PCR products is done using a specific hydrolysis probe labelled with fluorescent dyes (FAM as
reporter dye and a quencher) that binds the respective target sequence between the two primers (commonly
[5]
called “TaqMan chemistry” ).
5 Reagents and materials
Chemicals of recognized analytical grade, appropriate for molecular biology, shall be used. The water used
shall be double distilled or PCR grade water (i.e. nuclease and nucleic acid free). For all operations in which
gloves are used, it should be ensured that these are powder-free. The use of aerosol-protected pipette tips as
protection against cross contamination should be used.
5.1 Interlaboratory trial and control material.
5.1.1 DNA extracted from different salmon (Salmo salar) samples, taken in the context of official
control by the Institute for Hygiene and Environment in Hamburg, Germany for the construct-specific
method, or the National Institute for Health Sciences in Japan for the event-specific method.
5.1.2 Plasmid DNA, containing the target sequences of the AquAdvantage salmon construct-specific PCR
[1][2] [3]
methods , or event-specific PCR methods .
5.2 PCR reagents.
5.2.1 PCR master mix, contains thermostable DNA polymerase (for hot-start PCR), MgCl and
deoxyribonucleoside triphosphates (dNTPs).
5.2.2 TE buffer, c(Tris-HCl) = 1 mmol/l, c(EDTA) = 0,1 mmol/l (pH = 8,0).
5.2.3 Oligonucleotides (see Table 1).

ISO/TS 21569-10:2026(en)
Table 1 — Oligonucleotides
Final concentration
Name DNA sequence of oligonucleotide
in PCR
[1][2]
CS-GHc2 - T-AFP construct as the target sequence
GH-tFreeze-F 5′ - CTC CAC Agg TTT TgA CAT gTT CA - 3′ 340 nmol/l
GH-tFreeze-R 5′ - gCC AgC AAg AgC CCA TCT C - 3′ 340 nmol/l
GH-tFreeze-P 5′ -(FAM)- TTC CTA ATC TgT ATC Tgg gAA ACC gAA CCC T -(TAMRA)- 3′ 540 nmol/l
[3]
Atlantic salmon genome - P-AFP event as the target sequence
AquAd-F 5′ - TgC TgA TgC CTC TgA TAC CAC - 3′ 800 nmol/l
AquAd-R 5′ - ATg CCT CTA gTg CAA gTT CAg TC - 3′ 800 nmol/l
AquAd-P 5′ -(FAM)- CAg TAg TAC AAC gTT ggC AgA TgT ATg AgA ACT -(BHQ1)- 3′ 100 nmol/l
FAM: 6-Carboxyfluorescein; TAMRA: 5-Carboxytetramethylrhodamin; BHQ1: black hole quencher 1
Equivalent reporter dyes and/or (internal) quencher can be used for the probe if they can be shown to yield similar or better
results.
6 Apparatus
Requirements concerning apparatus and materials shall be in accordance with ISO 21569. The usual
laboratory apparatus and, in particular, the following shall be used.
6.1 Real-time PCR device, suitable for the excitation of fluorescent molecules and the detection of
fluorescence signals generated during PCR. In the interlaboratory trial, the real-time PCR devices listed in
Table 6 and 10 were used.
7 Procedure
7.1 Preparation of test samples
It should be ensured that the test sample used for DNA extraction is representative of the laboratory
sample (e.g. by grinding or homogenizing of the samples). Measures and operational steps to be taken into
consideration shall be as described in ISO 21571:2005 and ISO 24276.
7.2 DNA extraction
For the extraction of DNA from the test portion, the general instructions and requirements described in
ISO 21571 shall be followed.
7.3 PCR setup
The methods are described for a total volume of 25 μl per reaction mixture with the set-up given in Table 2.

ISO/TS 21569-10:2026(en)
Table 2 — Reaction set-up for PCR
Reagent Final concentration in 25 µl PCR reaction
Sample
...