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doi.org/10.6084/m9.figshare.1320632 and in the NCBI Bioproject PRJNA279807. 16S rRNA gene amplicons A 16S rRNA gene amplification step was done for DNA pools extracted from the seven microcosms targeted during the second sequencing phase as well as one microcosm representing the control condition. Primers pA (5�� AGA GTT TGA TCC TGG CTC AG 3��) and pH (5�� AAG GAG GTG ATC CAG CCG CA 3��) were used for this amplification. PCR products were then gel-purified (GE Healthcare illustra GFX PCR DNA and Gel Band Purification Kit) and cloned in a TOPO-TA using the TOPO-TA cloning kit for Sequencing (Invitrogen). 96 clones for each microcosm condition were randomly selected and inoculated in a 96 well plate. After TOPO-TA vector extraction, inserts were sequenced with Sanger Technology (Beckmann Coulter Genomics). Both primer M13for (5�� TGT AAA ACG ACG GCC AGT 3) and primer M13rev (5�� CAG GAA ROCK inhibition ACA GCT ATG ACC 3��) reactions were done. Sequences obtained with forward Non-specific serine/threonine protein kinase and reverse reactions were assembled and vector-purified with Seqman Software (Lasergen Seqman DNASTAR). Consensus sequences were then compiled by dataset for further data analysis. Taxonomical inference Genomic taxonomical affiliation was inferred using information extracted from (i) global sequence similarity with the RAST collection of genomes (Aziz et al., 2008), (ii) full or fragmented 16S rRNA genes present in assembled genomes, (iii) sequences related to 16S rRNA gene assembled after their screening at the genus level in unassembled metagenomic datasets, and (iv) 16S rRNA gene amplicon datasets. Results and discussion ESCs stimulate the reconstruction of genetic material from the soil microbiome We sequenced the genetic material of two microcosms for each ESC as well as the outlier microcosm replicate of the mercury enrichment #2 (see Figure S1), thus generating 23 metagenomic datasets. 1.05 (�� 0.17) million pyrosequencing reads of about 350 nucleotides were generated for each microcosm, representing a total of 24.05 million reads. We combined the 23 unassembled metagenomic dataset with the 13 Crizotinib cell line dataset previously generated from the natural community of the same site for community composition comparison purposes (Delmont et al., 2012). The latter dataset was generated with the same sequencing strategy and provides background regarding the natural and methodological fluctuation of the Park Grass soil microbiome. MG-RAST identified 4,081 genera and 8,541 distinct protein families (Pfams) from the combined datasets (Tables S1, S2). Among these, several taxonomical groups and Pfams varied significantly between different conditions (Kruskal-Wallis test, p-value