ISO 20397-1:2022

Overview

ISO 20397-1:2022 - Biotechnology - Massively parallel sequencing - Part 1: Nucleic acid and library preparation - specifies general requirements and provides guidance for evaluating nucleic acid sample quality and for preparing sequencing libraries prior to data generation. The standard focuses on upstream steps that directly affect massively parallel sequencing (MPS) results, including nucleic acid quantification, purity, integrity, library construction, and quality assessment.

Key Topics and Requirements

• Nucleic acid sample quality evaluation
  • Establish and document workflows for accurate, reproducible quantification.
  • Assess sample quantity, purity and integrity using validated methods.
  • Recommended analytical approaches include gel electrophoresis, capillary electrophoresis, microfluidic systems and PCR-based checks (as guidance, not prescriptive thresholds).
• Library preparation
  • General guidelines for fragmentation (mechanical, enzymatic, chemical), size distribution and purification.
  • Addition of universal sequences via end repair and adapter ligation, and strategies for barcoding/indexing multiple samples.
  • Size selection, amplification, purification/cleanup steps and library qualification.
• Library and sample quantification
  • Guidance on selecting appropriate quantification methods and developing QC procedures to ensure libraries are suitable for sequencing.
• Validation and controls
  • Use of positive, negative, no-template and spike-in controls; reference materials to monitor process performance.
• Contamination prevention
  • Requirements for primary sample evaluation, protocol design and operational procedures to minimize contamination risk.
• Supporting material
  • Informative annexes include a checklist for sample quality assessment, examples of platform-specific quality criteria, and a reference material list.

Applications and Who Uses It

ISO 20397-1:2022 is applicable to organizations performing MPS workflows where upstream sample handling and library prep influence downstream data quality:

• Molecular biology and genomics laboratories
• Clinical and diagnostic sequencing facilities
• Contract research organizations (CROs) and biotech companies
• Public health and research institutions performing high-throughput sequencing
• Laboratory managers and QA/QC personnel developing validated sequencing workflows

Adopting the standard helps improve reproducibility of sequencing data, reduce failed runs and contamination events, and supports compliance with laboratory quality systems.

Related Standards

• ISO 20395:2019 - Requirements for evaluating performance of nucleic acid quantification methods (qPCR/dPCR) - referenced for quantification performance.
• Other parts of the ISO 20397 series (see ISO website) address additional MPS topics and platform-specific considerations.

Keywords: ISO 20397-1:2022, massively parallel sequencing, MPS, nucleic acid quality, library preparation, library quality assessment, sequencing workflows, adapter ligation, barcoding/indexing, library quantification.