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CDS extraction and computation of codon adaptation index [31], as well as the analysis of codon usage at the second codon were carried out by using DAMBE (Xia 2001; Xia and Xie 2001).
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The preferred nucleotide usage at the third codon position of Toxocara mt protein-coding genes reflects the overall nucleotide composition of the mtDNA.
Furthermore, there is a significant wide variation in GC usage at the third synonymous position (GC3s ∼42.8%) (Table 1; Fig. 1).
Selection pressure on overall GC usage and nucleotide usage at the third codon position might not necessarily both act on the same viral segments (e.g. H1N1 PA and H1N1 HA).
An increase in A and decrease in G nucleotide usage at the third codon position with year of isolation was also observed in many of these viral genes (Additional File 12).
We found that average GC3 is significantly lower than GC1 and the observed GC (P-value < 0.05) which potentially explains that the genome composition is biased towards the dominating AT usage encoding genes at the third, 'wobble' position.
These changes reduced the overall GC content and the usage of G at the third codon position in the viral genome.
The study, the sixth in the series, also found that Netflix usage, at 76percentt of those surveyed, has surpassed combined cable and satellite usage for the first time.
The second point examined below is the preferential AA usage at the position surrounding 0 and -2.
I defended my usage at the time.
It wasn't in common usage at the time.
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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