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Concatenated sequences of these 175 genes were aligned using ClustalX v. 10.0.2 [ 40] or Muscle v. 3.6 [ 41] and Gblocks v. 0.91b [ 42] was applied to edit the alignment to remove gaps and poorly aligned regions.
Run UCHIME (Edgar et al., 2011) against the reference alignment to remove chimeric query sequences.
When this occurred in the internal part of the element, we manually edited the alignment to remove the columns corresponding to the repeat.
Our pipeline executes the following steps: Run UCHIME (Edgar et al., 2011 ) against the reference alignment to remove chimeric query sequences.
The other two alignments are matched negative controls generated by shuffling the order of positions in each observed alignment to remove all structural information.
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Before clustering, though, we filtered our set of TIC alignments to remove those that corresponded to intragenic alternate splicing events.
BS-Seeker2 circumvents these problems by using local alignments to remove the mismatched nucleotides from the end of the reads to maximize the mappability.
After trimming the multiple alignments to remove all noncoiled-coil domains, we developed a substitution model (which we named "CC") to describe the amino acid exchangeability of the coiled-coil domain, and compared this model with the LG, a general empirical model of protein evolution that was shown to outperform former general models in reconstruction of protein phylogenies (Le and Gascuel 2008).
Subsequently, these sequences were aligned using a global alignment algorithm to remove sequences for which the best blast hit represented only local homology.
I then manually trimmed the alignment edges to remove regions that were too divergent to provide phylogenetic signal.
(ii) We then perform local alignment (LA) to remove exons that are not highly conserved in sequence and exons that may cause ambiguity in RNA-seq read mapping.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com