Standard ISO standard · ISO 20395:2019

Biotechnology -- Requirements for evaluating the performance of quantification methods for nucleic acid target sequences -- qPCR and dPCR

Status: Valid

Buy this standard

Standard ISO standard · ISO 20395:2019

Biotechnology -- Requirements for evaluating the performance of quantification methods for nucleic acid target sequences -- qPCR and dPCR
Subscribe on standards - Read more Dölj
Price: 1 770 SEK
standard ikon pdf

PDF

Price: 1 770 SEK
standard ikon

Paper

Show more Show less
Scope
This document provides generic requirements for evaluating the performance and ensuring the quality of methods used for the quantification of specific nucleic acid sequences (targets).
This document is applicable to the quantification of DNA (deoxyribonucleic acid) and RNA (ribonucleic acid) target sequences using either digital (dPCR) or quantitative real-time PCR (qPCR) amplification technologies. It applies to target sequences present in nucleic acid molecules including double-stranded DNA (dsDNA) such as genomic DNA (gDNA) and plasmid DNA, single stranded DNA (ssDNA), complementary DNA (cDNA), and single stranded RNA (ssRNA) including ribosomal RNA (rRNA), messenger RNA (mRNA), and long and short non-coding RNA [microRNAs (miRNAs) and short interfering RNAs (siRNAs)], as well as double-stranded RNA (dsRNA).
This document applies to nucleic acids derived from biological sources such as viruses, prokaryotic and eukaryotic cells, cell-free biological fluids (e.g. plasma or cell media) or in vitro sources [e.g. oligonucleotides, synthetic gene constructs and in vitro transcribed (IVT) RNA].
This document is not applicable to quantification of very short DNA oligonucleotides (<50 bases).
This document covers:
— analytical design including quantification strategies (nucleic acid copy number quantification using a calibration curve as in qPCR or through molecular counting as in dPCR, quantification relative to an independent sample and ratio measurements) and use of controls;
— quantification of total nucleic acid mass concentration and quality control of a nucleic acid sample including assessment of nucleic acid quality (purity and integrity);
— PCR assay design, optimization, in silico and in vitro specificity testing;
— data quality control and analysis including acceptance criteria, threshold setting and normalization;
— method validation (precision, linearity, limit of quantification,...

Subjects

Biology, botany, zoology (07.080)


Buy this standard

Standard ISO standard · ISO 20395:2019

Biotechnology -- Requirements for evaluating the performance of quantification methods for nucleic acid target sequences -- qPCR and dPCR
Subscribe on standards - Read more Dölj
Price: 1 770 SEK
standard ikon pdf

PDF

Price: 1 770 SEK
standard ikon

Paper

Show more Show less

Product information

Language: English

Written by: ISO

International title:

Article no: STD-80014088

Edition: 1

Approved: 8/8/2019

No of pages: 50