National foreword

This British Standard is the UK implementation of ISO 20397‑1:2022.

The UK participation in its preparation was entrusted to Technical Committee BTI/1, Biotechnologies.

A list of organizations represented on this committee can be obtained on request to its committee manager.

Contractual and legal considerations

This publication has been prepared in good faith, however no representation, warranty, assurance or undertaking (express or implied) is or will be made, and no responsibility or liability is or will be accepted by BSI in relation to the adequacy, accuracy, completeness or reasonableness of this publication. All and any such responsibility and liability is expressly disclaimed to the full extent permitted by the law.

This publication is provided as is, and is to be used at the recipient’s own risk.

The recipient is advised to consider seeking professional guidance with respect to its use of this publication.

This publication is not intended to constitute a contract. Users are responsible for its correct application.

Compliance with a British Standard cannot confer immunity from legal obligations.

This British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 May 2022.

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 documents 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).

Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of any patent rights identified during the development of the document will be in the Introduction and/or on the ISO list of patent declarations received (see www.iso.org/patents).

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 276, Biotechnology.

A list of all parts in the ISO 20397 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.

Introduction

Massively parallel sequencing (MPS) is a high throughput analytical technology for nucleic acid sequencing. MPS methods can process thousands to billions of nucleotide sequence reads simultaneously in a single run, allowing whole genomes, transcriptomes and specific nucleic acid targets from different organisms to be analysed in a relatively short time.

MPS is used in many life science disciplines permitting determination and high throughput analysis of millions of nucleotide bases. The biological variability of deoxyribonucleic and ribonucleic acid polymers from living organisms provides challenges in accurately determining their sequences. The quality of sequence determination by MPS depends on many factors including, but not limited to, sample quality, library preparation, and sequencing data quality.

The quality of nucleic acids and libraries prepared for MPS is critical to obtaining high quality sequence data. Controlling the upstream processing steps of MPS and evaluating nucleic acid samples and libraries for their suitability for sequencing significantly improves MPS results, downstream analyses and ultimately conclusions dependent upon the MPS data.

1 Scope

This document specifies the general requirements for and gives guidance on quality assessments of nucleic acid samples. It specifies general guidelines for library preparations and library quality assessments prior to sequencing and data generation.

2 Normative references

3 Terms and definitions

4 Nucleic acid sample quality evaluation

4.1 General

4.2 Sample quantification

4.3 Sample purity

4.4 Sample integrity

5 Nucleic acid library preparation

5.1 General

5.2 Fragmentation

5.3 Addition of universal sequences

5.4 Size selection

5.5 Amplification

5.6 Purification and clean up procedures

5.7 Library quantification

5.8 Library qualification

6 Validation

7 Reference materials or controls

7.1 General

7.2 Control samples

7.3 Positive control

7.4 Negative control

7.5 No-template control

7.6 Spike-in control

7.7 Reference materials

8 Contaminations

8.1 General

8.2 Primary sample evaluation

8.3 Protocol and operation procedure

Bibliography

BS ISO 20397-1:2022

Biotechnology. Massively parallel sequencing - Nucleic acid and library preparation

Current

Published:

31 May 2022