title: DNA-seq aquatic fungi and bacteria from leaf litter
design: The DNA samples from all the replicates from each country were pooled for Illumina MiSeq sequencing using ITS primers for fungi (ITS3 F: 5'GCATCGATGAAGAACGCAGC 3' and ITS4R: 5'TCCTCCGCTTATTGATATGC3') and 16S primer pairs (515F 5'GTGCCAGCMGCCGCGGTAA3' and 806R 5'GGACTACHVGGGTWTCTAAT3') for bacteria (Bacteria and Archaea).The Illumina Paired-end read merger (PEAR) was used to merge the forward and reverse reads, followed by sorting the length of sequence reads from longest to shortest. The USEARCH algorithm was used to perform prefix dereplication and clustering at a 4% divergence. The clusters were then categorized into operational taxonomic units (OTUs) using the UPARSE OTU selection algorithm. UCHIME chimera detection software executed in de novo mode was used for chimera checking. Each of the original reads was then assigned back to their OTUs using the USEARCH global alignment algorithm. The centroid sequence from each cluster was run with USEARCH against a database of high quality 16S and ITS sequences from nr/nt. Each OTU was then assigned taxonomic information at 51 % conÞdence threshold.