ISO/PWI 26863

Biotechnology —Requirements and guidelines for absolute quantification of RNA-seq using calibrators traced to SI

Scope

This document describes a method for the absolute quantification of RNA-seq data through the utilization of spike-in reference materials (S-RMs) with metrological traceability to the International System of Units (SI).
This document provides technical guidance for:
a) The implementation of SI-traced S-RMs within RNA-seq library preparation and sequencing workflows;
b) The mathematical modeling and calibration procedures required to transform relative sequencing read counts into absolute transcript abundance;
c) The data analysis and validation steps necessary to ensure the consistency and comparability of absolute quantification results across different sequencing platforms and laboratories.
This document is intended for use in transcriptomic research, clinical diagnostics, and biotechnology applications where the absolute quantification of RNA transcripts is required.