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In order to assess topological support, 500 bootstrap replicates were performed with RAxML on each alignment with the parameters described above.
At least two runs of the M0 model were performed on each alignment to check the consistency of the log-likelihood values between the multiple runs.
In step 3, a sliding window analysis is carried out on each alignment to identify the best candidates i.e. with i. appropriate evolutionary rate for being potentially useful in phylogenetic analyses among close taxa and ii.
Independent Bayesian runs on each alignment yielded similar results.
Parameters of the evolution models for DNA and protein sequences were automatically estimated on each alignment using TREE-PUZZLE [ 47].
Bootstrap resampling was carried out 100 times on each alignment and the results were summarised with the majority-rule consensus method with a threshold of 70%.
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GENECONV was run using the default settings, which estimates p-values on 10,000 permutations of each alignment.
On the basis of each alignment, we partitioned substitutions by codon position and eliminated all non-LTR sequences with nonrandom distributions of substitutions across codon positions (χ test; P < 0.05).
The primers used for the qPCR were designed based on the consensus sequence of each alignment.
Alternatively, if the automatic selection of parameters options is selected, trimAl will compute specific score thresholds depending on the inherent characteristics of each alignment.
Two scores were calculated for each alignment, based on inclusion or exclusion of gaps in the alignment.
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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