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After a given round of alignment, those sequences that most obviously caused regions of problematic alignment were progressively removed until remaining sequences could be aligned without large indels.
SNPs missed by false alignments can be recovered by manually editing the problematic alignment.
Figure 1 illustrates the utility of uniqueome in identifying problematic alignment areas in an RNAseq dataset (Guttman et al., 2010).
Artefacts, for example deformation or compression of histological slices, and the problematic alignment of sections make 3D reconstructions a laborious and often highly subjective procedure.
In contrast, the tree topologies derived from the 12S and 16S rRNA genes (Figs. 6 and 7 in Holcroft [ 21]) were unorthodox, and she concluded that problematic alignment of the mitochondrial data could account for those results.
Also, our approach includes improved quality management, such as by automatically checking the GenBank sequences for strand polarity and annotation, by masking problematic alignment regions and by handling compositional heterogeneity [steps b.III, VII and XI].
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All problematic alignments were discarded, and the involved data clusters were sub-clustered using BLASTCLUST with a higher similarity threshold.
This portrait should be considered conservative because of the absence of some subtelomeric genes from our ortholog set, and because of the filters we applied, which removed orthologs with problematic alignments (see Materials and Methods).
Repeat regions are problematic for alignment algorithms and users tend to mask sequences before performing the alignments.
NCS are seen as problematic for alignment and phylogenetic analysis because of their structural constraints, non-randomness of evolution, and mutational changes such as slipped-strand mispairing, stem-loop secondary structure excision/repair, minute inversions, and intramolecular recombination [ 9].
Expecting the generally divergent nature of OBPs sequences (∼20% amino acid identity over all sequences) to make the sequence alignment problematic [49], we used several multiple sequence alignment (MSA) methods to evaluate potential different outcomes of using six alignment approaches (Table 2), which differ greatly in popularity and general approach to the MSA problem [50].
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Since I tried Ludwig back in 2017, I have been constantly using it in both editing and translation. Ever since, I suggest it to my translators at ProSciEditing.

Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com