Soil DNA Extraction Procedure Influences Protist 18S rRNA Gene Community Profiling Outcome

Susana S Santos, Inês Nunes, Tue Kjærgaard Nielsen, Samuel Jacquiod, Lars H Hansen, Anne Winding

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Abstract

Advances in sequencing technologies allow deeper studies of the soil protist diversity and function. However, little attention has been given to the impact of the chosen soil DNA extraction procedure to the overall results. We examined the effect of three acknowledged DNA recovery methods, two manual methods (ISOm-11063, GnS-GII) and one commercial kit (MoBio), on soil protist community structures obtained from different sites with different land uses. Results from 18S rRNA gene amplicon sequencing suggest that DNA extraction method significantly affect the replicate homogeneity, the total number of operational taxonomic units (OTUs) recovered and the overall taxonomic structure and diversity of soil protist communities. However, DNA extraction effects did not overwhelm the natural variation among samples, as the community data still strongly grouped by geographical location. The commercial DNA extraction kit was associated with the highest diversity estimates and with a corresponding higher retrieval of Excavata, Cercozoa and Amoebozoa-related taxa. Overall, our findings indicate that this extraction offers a compromise between rare and dominant taxa representation, while providing high replication reproducibility. A comprehensive understanding of the DNA extraction techniques impact on soil protist diversity can enable more accurate diversity assays.

Original languageEnglish
JournalProtist
Volume168
Issue3
Pages (from-to)283-293
Number of pages11
ISSN1434-4610
DOIs
Publication statusPublished - 2017

Keywords

  • Agriculture
  • GnS-GII
  • ISOm
  • MoBio
  • protist diversity
  • taxonomic structure.

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