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Supplemental Information

16S fusion primers used in this study

Figure S1. 16S Fusion primers developed in this study. They include flow cell and sequencing primer binding regions for current Illumina sequencers. The amplified fragment has a size fo ~157 bp and can be sequenced directly after purification (one step PCR). Up to 10 samples can be uniquely tagged from forward and reverse direction and pooled in one NextSeq run. The bases used for shifting on Ins_F and Ins_R can be used to uniquely tag samples (inline barcodes). It is recommended that all 10 primer pairs are used in the following combination to maximize sequence diversity and reduce effects of tag switching by uniquely tagging samples from both sides: P5_Ins_R0+P7_Ins_F4, P5_Ins_R1+P7_Ins_F3, P5_Ins_R2+P7_Ins_F2, P5_Ins_R3+P7_Ins_F1, P5_Ins_R4+P7_Ins_F0, P5_Ins_F0+P7_Ins_R4, P5_Ins_F1+P7_Ins_R3, P5_Ins_F2+P7_Ins_R2, P5_Ins_F3+P7_Ins_R1, P5_Ins_F4+P7_Ins_R0

DOI: 10.7287/peerj.preprints.1855v1/supp-1

Distribution of reads obtained by NextSeq and number of reads discarded throughout the different bioinformatics processing steps

Figure S2. Number of sequences obtained per sample after library demultiplexing (A) and percentage of sequences excluded in different bioinformatic analysis steps (B). A: Library demultiplexing; Numbers above bars indicate the relative contribution (in percent) to the total number of sequences obtained for each sample. Sequencing started with Ins_F (white) or Ins_R (black) is indicated by bar color. B: Number of reads excluded in data processing steps. Mean percentage of sequence abundance in each processing step is written in brackets. Ins_F / Ins_R primer bias was tested with a t-test.

DOI: 10.7287/peerj.preprints.1855v1/supp-2

Sequence of each OTU with abundance of assigned reads and assigned taxonomy

DOI: 10.7287/peerj.preprints.1855v1/supp-3

Distribution of OTUs across the 52 taxa

DOI: 10.7287/peerj.preprints.1855v1/supp-4

Raw number of reads assigned to each of the 52 taxa for 16S and COI across the 10 replicates

DOI: 10.7287/peerj.preprints.1855v1/supp-5

R scripts used in this study to process sequence data and create plots

DOI: 10.7287/peerj.preprints.1855v1/supp-6

Additional Information

Competing Interests

The authors declare that they have no competing interests.

Author Contributions

Vasco Elbrecht conceived and designed the experiments, performed the experiments, analyzed the data, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.

Pierre Taberlet analyzed the data, wrote the paper, reviewed drafts of the paper, designed the ins 16 primers.

Tony Dejean designed the ins 16 primers.

Alice Valentini designed the ins 16 primers.

Philippe Usseglio-polatera designed the ins 16 primers.

Jean-nicolas Beisel designed the ins 16 primers.

Eric Coissac analyzed the data, reviewed drafts of the paper, designed the ins 16 primers.

Frederic Boyer designed the ins 16 primers.

Florian Leese conceived and designed the experiments, performed the experiments, wrote the paper, reviewed drafts of the paper.

DNA Deposition

The following information was supplied regarding the deposition of DNA sequences:

BOLDsystems (16S Sanger data): DS-TMIX16S

SRA (Illumina data): SRR2217415

Data Deposition

The following information was supplied regarding data availability:

Additional raw data has been supplied as a Supplemental Dataset

Funding

Florian Leese and Vasco Elbrecht are supported by a grant of the Kurt Eberhard Bode foundation to Florian Leese. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.


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