title: An Illumina metabarcoding pipeline for fungi.
abstract: High-throughput metabarcoding studies on fungi and other eukaryotic microorganisms are rapidly becoming more frequent and more complex, requiring researchers to handle ever increasing amounts of raw sequence data. Here, we provide a flexible pipeline for pruning and analyzing fungal barcode (ITS rDNA) data generated as paired-end reads on Illumina MiSeq sequencers. The pipeline presented includes specific steps fine-tuned for ITS, that are mostly missing from pipelines developed for prokaryotes. It (1) employs state of the art programs and follows best practices in fungal high-throughput metabarcoding; (2) consists of modules and scripts easily modifiable by the user to ensure maximum flexibility with regard to specific needs of a project or future methodological developments; and (3) is straightforward to use, also in classroom settings. We provide detailed descriptions and revision techniques for each step, thus giving the user maximum control over data treatment and avoiding a black-box approach. Employing this pipeline will improve and speed up the tedious and error-prone process of cleaning fungal Illumina metabarcoding data.
authors: Biodiversity and Climate Research Centre, Senckenberg Gesellschaft für Naturforschung Senckenberganlage 25, 60325, Frankfurt/Main, Germany.
authors: Miklós
authors: M
authors: Bálint
authors: Biodiversity and Climate Research Centre, Senckenberg Gesellschaft für Naturforschung Senckenberganlage 25, 60325, Frankfurt/Main, Germany ; Institut für Ökologie, Evolution und Diversität, Goethe Universität Frankfurt Max-von-Laue-Str. 13, 60438, Frankfurt/Main, Germany.
authors: Philipp-André
authors: PA
authors: Schmidt
authors: Institut für Ökologie, Evolution und Diversität, Goethe Universität Frankfurt Max-von-Laue-Str. 13, 60438, Frankfurt/Main, Germany.
authors: Rahul
authors: R
authors: Sharma
authors: Biodiversity and Climate Research Centre, Senckenberg Gesellschaft für Naturforschung Senckenberganlage 25, 60325, Frankfurt/Main, Germany ; Institut für Ökologie, Evolution und Diversität, Goethe Universität Frankfurt Max-von-Laue-Str. 13, 60438, Frankfurt/Main, Germany.
authors: Marco
authors: M
authors: Thines
authors: Biodiversity and Climate Research Centre, Senckenberg Gesellschaft für Naturforschung Senckenberganlage 25, 60325, Frankfurt/Main, Germany ; Institut für Ökologie, Evolution und Diversität, Goethe Universität Frankfurt Max-von-Laue-Str. 13, 60438, Frankfurt/Main, Germany.
authors: Imke
authors: I
authors: Schmitt
keywords: community ecology; data pruning; high throughput; internal transcribed spacer rDNA; next-generation sequencing