| Summary: | A pilot run was carried out on Antarctic microbial mat samples in order to test different protocols for 454 pyrosequencing and bioinformatic analyses. An artificial community was assembled using22 cyanobacterial strains in two different abundance distributions. Bioinformatic analyses were carried out on the artificial communities using variations of the mothur and UPARSE pipelines. Moreover, different DNA extraction methods, technical replicates and biases arising from direct barcoded primer amplification were assessed using five microbial mat and soil crust samples. A great variation in richness estimates was observed using different bioinformatic pipelines, with UPARSE generating the most consistent results. On the other hand, community structures observed using different DNA extraction protocols were statistically similar. The same was observed for the technical replicates. Moreover, the use of barcoded primers did not influence the observed community structure. CCAMBIO
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