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These n-grams were evaluated in relation to their relevance and manually refined by translation students with knowledge of the domain.
Genes were predicted on the assembled genome sequence using the FgenesB program (Softberry, Inc .. Automated FgenesB annotations were manually refined by reviewing Reciprocal Best Hits (RBH) to Nitrosopumilus maritimus and Cenarchaeum symbiosum A and BLAST searches against the GenBank database.
Annotations were manually refined by comparison with genes of more closely related species.
The CGOB pillar dataset was manually refined by examining regions of dubious synteny and singleton genes.
Finally, the automatically generated clusters were manually refined by a graphical cluster cutting program.
Protein sequences were aligned with MUSCLE [ 36] and manually refined by removing sites at which all sequences except one or two have alignment gaps.
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The putative ancestral sequences were subsequently inspected by eye and manually refined.
For each dataset, proteins have been aligned by ClustalW [ 62], manually refined using ED [ 63], and then concatenated into a super-matrix using SCaFoS [ 64].
Alignment quality was manually refined in the SeqLab (GCG, Wisconsin package) editor by moving residues.
Sequences of each of the 11 ribosomal protein genes were aligned individually using default settings in Muscle 3.8 (Edgar, 2004) and were realigned by predicted translation; alignments were manually refined and stop-codon sites removed.
The sequences were aligned by Clustal X 2.0 [ 47], manually refined and 82 amino acid residues corresponding to the YGGT domain were used for the phylogenetic analyses.
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