Abstract


IN NEXT GENERATION AMPLICON SEQUENCING PROJECTS IS REPLICATION A NECESSARY EXPENSE?

Environmental samples are being collected and sequenced around the globe with one of the main aims being the characterisation and comparison of bacterial community diversity. Experimental design is limited by cost and bioinformatics techniques can be selected for convenience and/or to facilitate comparisons with other studies. Here we consider questions of experimental design for 16S rRNA gene sequencing projects and the implications for downstream analyses using a South Indian Ocean marine sample. Specifically we address issues of replication and robustness using triplicate independent PCRs and performing comparative analyses including rarefaction. Combining the reads across all three PCRs, we see that saturation of the rarefaction curve begins to occur at 7 million reads. We also show that the taxonomic assignments for the OTUs display some differences between Silva 115 reference set and the Greengenes databases, yet overall, the taxonomic composition of the samples to family level are substantially the same. The dominance of Cyanobacteria (Prochlorococcus), and the bacterial diversity observed in this study is consistent with other open ocean studies in the same region.

Authors

Schroeder, J. L., Marine Biological Association of the UK, United Kingdom, joasch@mba.ac.uk

Balestreri, C., Marine Biological Association of the UK, United Kingdom, cecbal@mba.ac.uk

Yilmaz, P., Max Plank Institute for Marine Microbiology, Germany, pyilmaz@mpi-bremen.de

Paszkiewicz, K. H., Biosciences, Exeter Sequencing Service, United Kingdom, K.H.Paszkiewicz@exeter.ac.uk

Moore, K. A., Biosciences, Exeter Sequencing Service, United Kingdom, K.A.Moore@exeter.ac.uk

Thorpe, S. E., British Antarctic Survey, United Kingdom, seth@bas.ac.uk

Glockner, F. O., Max Plank Institute for Marine Microbiology, Germany, fgloeckn@mpi-bremen.de

Schroeder, D., Marine Biological Association of the UK, United Kingdom, dsch@mba.ac.uk

Details

Oral presentation

Session #:075
Date: 2/26/2015
Time: 18:00
Location: Andalucia 1 (Floor 1)

Presentation is given by student: No