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A hyper-variable loop portion of the 16S alignment could not be aligned with confidence, so nine internal positions were excluded.
Regions that could not be aligned with confidence were omitted from the phylogenetic analyses [the final alignment is given in Additional file 10].
They were not included in Fig. 1 because their sequences could not be aligned with confidence.
Nonhomologous insertions and sequence characters that could not be aligned with confidence were removed manually.
Nonhomologous insertions and sequence characters that could not be aligned with confidence were manually removed.
Typically, phylogenetic reconstruction is suited for analyzing subsets of homologous genes that can be aligned with confidence.
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Sequences were assembled with Sequencer 4.5 (GeneCode Corporation, Ann Arbor, MI, USA) and aligned by eye using BIOEDIT version 7.0.5.3 [16]; we observed no indels in this coding region of the mitochondrial genome and therefore all base positions were aligned with confidence in positional homology.
Only sequence positions that are aligned with confidence are retained for model construction.
Moreover, Golubchik et al. ([3]) showed that the absence of amino acid residues often leads to an incorrect placement of gaps in the alignments, even when the sequences were otherwise identical, and, for a given alignment, not all amino acid positions will be aligned with equal confidence [4].
Measuring positive diversifying selection is only possible in families where the coding sequences can be aligned with sufficient confidence and many of the CSEPs have diverged so much that they are too different for a reliable analysis.
For identification, we used only those features that could be aligned with the storage phosphor images and selected from the gel with high confidence.
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