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We used the following parameters: parsing type none, spatial, valid, gain and temporal calibration automatic, temporal algorithm sequence, temporal valid uncertainty false, alignment uncertainty 15, calibration frequency 15, and video model general model.
Forward and reverse reads for all regions were assembled and aligned using Geneious Pro (Biomatters, New Zealand), with the alignments being subsequently manually edited in BioEdit 7.1.3 (Hall 1999); inversions were offset and the final alignment was 6905 bases long, with 39 bases excluded due to alignment uncertainty.
An alternative, more statistically justified approach to assess alignment uncertainty is the use of probabilistic evolutionary models for joint estimation of phylogeny and alignment.
The HoT (Heads-Or-Tails) score can be used as a measure of site-specific alignment uncertainty due to the existence of multiple co-optimal solutions.
Alignment uncertainty can cause discrepancies in phylogenetic inference [35], and possibly in other bioinformatic analyses.
Additionally, they show that it is possible to make inferences despite considerable alignment uncertainty.
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A 10 bp overlap between the partner sequences was allowed at the fusion point to account for alignment uncertainties.
Alignment uncertainties were removed using trimAl (automated1 mode) (Capella-Gutierrez et al. 2009).
This latter was checked and edited by hand to remove alignment uncertainties.
This latter was checked and edited by TrimAl to remove alignment uncertainties (http://phylemon.bioinfo.cipf.es/utilities.html, last accessed October 31, 2014) (Capella-Gutierrez et al. 2009).
The GUIDANCE program calculates a similar site-specific confidence measure based on the robustness of the alignment to uncertainty in the guide tree that is used in progressive alignment programs.
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