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[12] have recently presented a methodology for de novo ncRNA annotation which relies on an input multiple alignment only for homology detection, and so is capable of detecting conserved structure even in the presence of local mis-alignment.
These were concatenated in a new multiple alignment only containing third bases.
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In these cases we did not do any filtering of sequences in multiple alignments but we only weighted sequences using the position-based method of sequence weighing [ 12].
The question of reconstructing phylogenies directly without multiple alignment has only recently been tackled [20] with promising results.
Consider a multiple alignment of only three reads { A, B, C}, where A conflicts with B, B conflicts with C, but A and C do not overlap.
For each such cluster, multiple alignments including only one paralog at a time were constructed using Clustal Omega (Sievers et al. 2011) and trimmed using Aliscore and Alicut (Kück 2009; Misof and Misof 2009).
It is easy to see that the resulting multiple alignment would not only be biologically questionable, but it would also obtain a numerically lower score as it would involve only two pairwise alignments of the motif.
According to the multiple alignment, we found only two conserved regions for the PCR-primers.
Similar promoters with unambiguous 5'-UTR multiple alignment are found only in selected species of Brassicaceae: Arabis hirsuta, Barbarea verna, Crucihimalaya wallichii, Draba nemorosa, Lepidium virginicum, Lobularia maritima, Nasturtium officinale and Olimarabidopsis pumila.
Examination of sequences in the multiple alignment shows not only that Cys tends to occur every third residue in the carboxyl-terminal region, but also that differences between closely related sequences commonly feature tripeptide indels.
Only multiple alignment positions where the sequence coverage exceeded 6× were used in order to avoid low quality regions.
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