prEN ISO 17601

SLOVENSKI STANDARD
01-oktober-2024
Kakovost tal - Ocena številčnosti izbranih sekvenc mikrobnih genov s
kvantitativnim PCR analizatorjem v talnih ekstraktih DNK (ISO/DIS 17601:2024)
Soil quality - Estimation of abundance of selected microbial gene sequences by
quantitative PCR from DNA directly extracted from soil (ISO/DIS 17601:2024)
Bodenbeschaffenheit- Ermittlung der Häufigkeit ausgewählter mikrobieller
Gensequenzen durch quantitative PCR aus DNA-Boden-Extrakten (ISO/DIS
17601:2024)
Qualité du sol - Estimation de l'abondance de séquences de gènes microbiens par
amplification par réaction de polymérisation en chaîne (PCR) quantitative à partir d'ADN
directement extrait du sol (ISO/DIS 17601:2024)
Ta slovenski standard je istoveten z: prEN ISO 17601
ICS:
13.080.30 Biološke lastnosti tal Biological properties of soils
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

DRAFT
International
Standard
ISO/DIS 17601
ISO/TC 190/SC 4
Soil quality — Estimation of
Secretariat: AFNOR
abundance of selected microbial
Voting begins on:
gene sequences by quantitative PCR
2024-08-15
from DNA directly extracted from
Voting terminates on:
soil
2024-11-07
Qualité du sol — Estimation de l'abondance de séquences
de gènes microbiens par amplification par réaction de
polymérisation en chaîne (PCR) quantitative à partir d'ADN
directement extrait du sol
ICS: 13.080.30
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENTS AND APPROVAL. IT
IS THEREFORE SUBJECT TO CHANGE
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Reference number
ISO/DIS 17601:2024(en)
DRAFT
ISO/DIS 17601:2024(en)
International
Standard
ISO/DIS 17601
ISO/TC 190/SC 4
Soil quality — Estimation of
Secretariat: AFNOR
abundance of selected microbial
Voting begins on:
gene sequences by quantitative PCR
from DNA directly extracted from soil
Voting terminates on:
Qualité du sol — Estimation de l'abondance de séquences
de gènes microbiens par amplification par réaction de
polymérisation en chaîne (PCR) quantitative à partir d'ADN
directement extrait du sol
ICS: 13.080.30
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENTS AND APPROVAL. IT
IS THEREFORE SUBJECT TO CHANGE
AND MAY NOT BE REFERRED TO AS AN
INTERNATIONAL STANDARD UNTIL
PUBLISHED AS SUCH.
This document is circulated as received from the committee secretariat.
IN ADDITION TO THEIR EVALUATION AS
BEING ACCEPTABLE FOR INDUSTRIAL,
© ISO 2024
TECHNOLOGICAL, COMMERCIAL AND
USER PURPOSES, DRAFT INTERNATIONAL
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
STANDARDS MAY ON OCCASION HAVE TO
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BE CONSIDERED IN THE LIGHT OF THEIR
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Published in Switzerland Reference number
ISO/DIS 17601:2024(en)
ii
ISO/DIS 17601:2024(en)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 2
5 Test materials . 4
5.1 DNA .4
5.2 Bacteria .4
5.3 Plasmid .4
5.4 Enzyme .4
5.5 Chemicals .4
5.6 Product for bacterial culture medium .5
5.7 Buffer and reagents .5
6 Apparatus . 6
7 Procedure . 6
7.1 qPCR standard preparation and calibration of qPCR assay (task 1) .6
7.1.1 General .6
7.1.2 Amplicon design (task 1, step 1) .6
7.1.3 qPCR standard preparation (task 1, step 2) .7
7.1.4 Isolate DNA, environmental DNA, artificial DNA .7
7.1.5 Calibration of the qPCR (task 1, step 3) .9
7.2 Preparation of soil DNA template and inhibition test (task 2) .10
7.2.1 General .10
7.2.2 Soil DNA preparation (task 2, step 4) . .10
7.2.3 Inhibition test (task 2, step 5) .10
7.2.4 Dilution of DNA template . 12
7.3 qPCR assay (task 3) . . . 12
7.3.1 General . 12
7.3.2 qPCR . . 12
7.4 Validation and analysis of qPCR assay (task 4) . 12
7.4.1 General . 12
7.4.2 Validation of the qPCR assay . 12
7.4.3 Calculation of the copy number of the gene of interest in the soil DNA extract . 13
8 Examination of the critical steps of the qPCR assay .13
9 Expression of the results of the qPCR assay . 14
10 International ring test . 14
11 Test report . 14 ®
Annex A (informative) Description of principal steps of TaqMan qPCR assay .15
Annex B (informative) International ring-test for evaluating qPCR to quantify the abundance
of selected microbial gene sequences from DNA directly extracted from soil. 17
Annex C (informative) Examples of well established primer systems for a qPCR based
quantification of marker genes in soil samples .30
Bibliography .33

iii
ISO/DIS 17601:2024(en)
Foreword
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