Did you evaluate other methods for assignnment of taxonomy other than BLAST?
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In your conclusion for the optimized steps, number 7 recomends assigning taxonomy with BLAST. Did you test other approaches to taxonomy assignment?
example: Wang Q, Garrity GM, Tiedje JM, Cole JR (2007). Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73: 5261-7.
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