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The default sequences of the target candidates in the server are of old version, hence the updated EST sequences of P. vulgaris from NCBI GenBank were used as target candidates.
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That could be the default sequence, displayed on the default table of contents.
Each segment was aligned against 7SK using the RNA structure alignment tool Foldalign [72] with the maximum motif length set to 100 and default sequence length difference settings.
Aux/IAA7 (IAA7) was the default sequence unless specified otherwise (Table 1).
Data was processed by using the default sequence analysis parameters and analyzed with Beckman Coulter CEQ 8000 software.
The sequence selected in the first run for each OTU becomes the default sequence for a given OTU.
For each protein, a large number of alternative MSAs was randomly generated from a default sequence alignment (Fig. 1).
As long as the list of complete sequences included in the OTU remains unchanged (i.e. no sequence are added or removed), SCaFoS automatically keeps the default sequence.
All subsequent analyses were performed for default sequence alignment settings, in order to be able to compare findings reported here with previous studies [ 38, 42].
We used PRANK+F with codon option [ 71] for creating the alignments and GUIDANCE [ 42] with the default sequence quality cut-off and a column cut-off of 0.97 to remove problematic sequences and unreliable sites from the initial alignments.
Strong, pre-existing cryptic sites outside of the default sequence analysis window (54 nt circumscribing the mutation) have also been missed because the window was not expanded to include these sites.
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