ITS (internal transcribed spacer) amplicons were generated with 35 cycles using Invitrogen AccuPrime PCR reagents. These amplicons were used in a second PCR reaction, applying Illumina Nextera XT v.2 (Illumina) barcoded primers to uniquely index each sample as previously described20 (link). All libraries were processed for quality control using the DNA 1000 Bioanalyzer (Agilent) and Qubit (Life Technologies) to validate and quantify library construction before preparing a paired-end flow cell. The samples were randomly divided among flow cells to minimize sequencing bias. Clonal bridge amplification (Illumina) was performed using a cBot (Illumina); 2× 250-base pair (bp) sequencing-by-synthesis was performed using the Illumina MiSeq platform.
Other organizations :
Kiel University, Cornell University, Humboldt-Universität zu Berlin, Charité - Universitätsmedizin Berlin, Clinical Research Center Kiel, University of Lübeck, Leibniz-Institut für Naturstoff-Forschung und Infektionsbiologie e. V. - Hans-Knöll-Institut (HKI), Friedrich Schiller University Jena
Bioanalyzer and Qubit for library quality control and quantification
Clonal bridge amplification (Illumina) using a cBot
Illumina MiSeq platform for sequencing
controls
No positive or negative controls were explicitly mentioned.
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