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We used this approach of determining optimal DNA substitution model, testing for recombination, and using the resulting DNA substitution model and partitioned dataset (when recombination was detected) as inputs for the four codon based tests of selection.
Thus, rather than picking among these methods we cautiously employed several codon based tests of selection which covered the range of fundamental methodological assumptions, presented a full disclosure of their findings, and identified only codons which found some support across these methods (Table 5).
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We used the H statistic developed in Fay and Wu [ 14] as a frequency based test of selection from species polymorphism.
Our second test was a codon based test of selection that looked for both positive (diversifying) and negative (purifying) selection while making no assumptions regarding demographic history and incorporating recombination effects [ 47].
Haplotype based tests of directional selection on malaria parasites in West Africa have previously identified selection occurring over very recent time frames, with strong signatures linked to the use of chloroquine and the antifolate drugs sulphadoxine-pyrimethamine as first line malaria therapies [ 17, 19, 22].
This topology was used in subsequent codon-based tests of selection.
A phylogenetic tree is required for the site-based tests of selection implemented in PAML.
To test between these alternatives, we performed codon-based tests of selection using orthologs from eight Caenorhabditis species.
This rate (128) is high enough to cause false positives in site-based tests of selection, especially the M7 vs. M8 test [ 65].
We next used codon- and branch-based tests of selection to test for positive selection on the R&R Consensus of An. gambiae CPRs.
As we showed in Results/Discussion, part 3.1, linkage of whole clonal genomes prevents standard multilocus-based tests of selection and indirect tests are necessary.
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