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Second, aligning divergent sequences is difficult, and the alignment itself can create a strong phylogenetic bias.
This however is difficult without the information from the alignment itself.
In other words, the alignment itself has little effect on the homology estimation.
Most importantly, we confirmed that our method neither depends on the alignment itself, nor on the level of sequence identity.
This can be interpreted as additional evidence beyond the alignment itself, making it more likely that the mammalian hits are indeed homologs.
The Fhom Estimator, which is independent of the alignment itself, enables estimating the amount of noise added, and is therefore beneficial for the E-Value threshold fine tuning.
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The final score (in the bottom right cell) is then the score of the best possible alignment between the two sequences; that alignment itself is given by the path or paths that yield that score.
Besides the potential problem of limited taxa and gene sampling, the tree topology may be more influenced by the final alignment itself than by the phylogenetic reconstruction method [ 54, 55].
In fact, as was demonstrated previously, the cell alignment itself requires 18 24 h of exposure to flow, but the cells respond already within first hours by cytoplasmic actin-reorientation (Osborn et al., 2006).
Protein secondary structure was predicted using a multiple alignment as the input for the JPRED program [ 48], with information extracted from a PSSM, HMM, and the seed alignment itself.
Another reason could be the incompleteness of the multi-species alignment itself.
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