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BS EN ISO 17601:2018

BS EN ISO 17601:2018

$167.15

Soil quality. Estimation of abundance of selected microbial gene sequences by quantitative PCR from DNA directly extracted from soil

Published By Publication Date Number of Pages
BSI 2018 40
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This International Standard specifies the crucial steps of a quantitative real-time polymerase chain reaction (qPCR) method to measure the abundance of selected microbial gene sequences from soil DNA extract which provides an estimation of selected microbial groups.

It is noteworthy that the number of genes is not necessarily directly linked to the number of organisms that are measured. For example, the number of ribosomal operon is ranging from one copy to 20 copies in different bacterial phyla. Therefore, the number of 16S rRNA sequences quantified from soil DNA extracts does not give an exact estimate of the number of soil bacteria. Furthermore, the number of sequences is not necessarily linked to living microorganisms and can comprise sequences amplified from dead microorganisms.

PDF Catalog

PDF Pages PDF Title
2 undefined
4 European foreword
Endorsement notice
6 Foreword
7 Introduction
8 1 Scope
2 Normative references
3 Terms and definitions
9 4 Principle
11 5 Test materials
5.1 DNA
5.2 Bacteria
5.3 Plasmid
5.4 Enzyme
5.5 Chemicals
12 5.6 Product for bacterial culture medium
5.7 Buffer and reagents
13 6 Apparatus
7 Procedure
7.1 qPCR standard preparation and calibration of qPCR assay (task 1)
7.1.1 General
7.1.2 Amplicon design (task 1, step 1)
14 7.1.3 qPCR standard preparation (task 1, step 2)
7.1.4 Isolate DNA, environmental DNA, artificial DNA
16 7.1.5 Calibration of the qPCR (task 1, step 3)
17 7.2 Preparation of soil DNA template and inhibition test (task 2)
7.2.1 General
7.2.2 Soil DNA preparation (task 2, step 4)
7.2.3 Inhibition test (task 2, step 5)
19 7.2.4 Dilution of DNA template
7.3 qPCR assay (task 3)
7.3.1 General
7.3.2 qPCR
7.4 Validation and analysis of qPCR assay (task 4)
7.4.1 General
7.4.2 Validation of the qPCR assay
20 7.4.3 Calculation of the copy number of the gene of interest in the soil DNA extract
21 8 Examination of the critical steps of the qPCR assay
9 Expression of the results of the qPCR assay
10 International ring test
11 Test report
22 Annex A (informative) Description of principal steps of TaqManĀ® qPCR assay
24 Annex B (informative) International ring-test for evaluating qPCR to quantify the abundance of selected microbial gene sequences from DNA directly extracted from soil
37 Bibliography

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BS EN ISO 17601:2018
$167.15