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The edges shown in this figure represent a mean pairwise alignment score for all-by-all comparisons of these 4298 sequences that is better than an E-value cutoff of 1 × 10 80, with a median percent identity of 43% and a median alignment length of 377 residues.
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Table 5 shows the mean pairwise percentage of alignment sites with a gap, calculated separately for comparisons within Drosophila, within Sophophora, and between the two clades.
Sequence similarity in this data set ranges from 47% to 99% mean pairwise identity in alignments of 40 nt to 400 nt length and of 2 to 6 sequences.
Consequently, studies using methods slightly different in their mathematical parameterisation have been dramatically affected by the properties of MSAs (i.e., Number of sequences in the alignment, mean pairwise sequence amino acid distances, etc).
The extent to which sequences are diverse and contain covarying mutations, as measured by the mutual information [ 9], entropy [ 23] of base-pairing regions, and the mean pairwise sequence identity between alignment sequences.
All structures exhibit very similar conformations of the protein with a mean all-atom Root-Mean-Square Deviation (RMSD) for pairwise alignment of 1.0 Å, calculated in PyMOL using the align command with the cycles flag set to zero [28].
Based on the mean pairwise estimates on single alignments, the smallest average Ks value was found for the AB/OV comparison (0.0289), followed by similarly low values between tilapia and both OV and AB (0.0732 and 0.0746) and between medaka and all other species comparisons (0.8657 0.8695).
The remaining 188 alignments were realigned and all which did not meet the prerequisites of RNAstrand were discarded: 15 alignments were shorter than the minimum length for which RNAstrand was trained with, 5 alignments had a mean pairwise identity smaller than 50%, and one alignment contained of too many gaps.
Alignments containing more than 6 sequences were reduced to 6 sequences by rnazWindow.pl which optimizes the final alignment for a mean pairwise identity.
2 families had no sampled alignments with a mean pairwise sequence identity larger than 60%.
To summarize the overall diversity of these repertoires, we calculated the proportion of segregating sites (%S) and the mean pairwise diversity of each of the 12 nucleotide alignments and 9 amino acid alignments.
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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