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Petric, I.* ; Philippot, L.* ; Abbate, C.* ; Bispo, A.* ; Chesnot, T.* ; Hallin, S.* ; Laval, K.* ; Lebeau, T.* ; Lemanceau, P.* ; Leyval, C.* ; Lindström, K.* ; Pandard, P.* ; Romero, E.* ; Sarr, A.* ; Schloter, M. ; Simonet, P.* ; Smalla, K.* ; Wilke, B.M.* ; Martin-Laurent, F.

Inter-laboratory evaluation of the ISO standard 11063 "Soil quality - Method to directly extract DNA from soil samples".

J. Microbiol. Methods 84, 454-460 (2011)
DOI PMC
Open Access Green as soon as Postprint is submitted to ZB.
Extracting DNA directly from micro-organisms living in soil is a crucial step for the molecular analysis of soil microbial communities. However, the use of a plethora of different soil DNA extraction protocols, each with its own bias, makes accurate data comparison difficult. To overcome this problem, a method for soil DNA extraction was proposed to the International Organization for Standardization (ISO) in 2006. This method was evaluated by 13 independent European laboratories actively participating in national and international ring tests. The reproducibility of the standardized method for molecular analyses was evaluated by comparing the amount of DNA extracted, as well as the abundance and genetic structure of the total bacterial community in the DNA extracted from 12 different soils by the 13 laboratories. High quality DNA was successfully extracted from all 12 soils, despite different physical and chemical characteristics and a range of origins from arable soils, through forests to industrial sites. Quantification of the 16S rRNA gene abundances by real time PCR and analysis of the total bacterial community structure by automated ribosomal intergenic spacer analysis (A-RISA) showed acceptable to good levels of reproducibility. Based on the results of both ring-tests, the method was unanimously approved by the ISO as an international standard method and the normative protocol will now be disseminated within the scientific community. Standardization of a soil DNA extraction method will improve data comparison, facilitating our understanding of soil microbial diversity and soil quality monitoring.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Soil DNA extraction; Standardization; Inter-laboratory assay; DNA fingerprint; qPCR
ISSN (print) / ISBN 0167-7012
e-ISSN 1872-8359
Journal Journal of Microbiological Methods
Quellenangaben Volume: 84, Issue: 3, Pages: 454-460 Article Number: , Supplement: ,
Publisher Elsevier
Publishing Place Amsterdam
Reviewing status Peer reviewed
Institute(s) Comparative Microbiome Analysis (COMI)
Institute of Soil Ecology (IBOE)