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To test directly for epistatic interactions between the detected QTLs, we conducted an ANOVA using the pairwise epistasis model.
Using the pairwise epistasis model analysis separately for all metabolites, we identified all significant main effect markers, individual genetic loci, and interactions, epistatic terms.
For each metabolite, open diamonds indicate the center of phenotype estimated using the pairwise epistasis model while closed diamonds indicate the estimated location using the three-way epistasis model.
Dotted lines show estimated variance using solely the main effect loci without interactions (additive model), solid lines show the distribution of estimated variance across the metabolites using the pairwise epistasis model while the dashed lines are the results for the three-way epistasis model including the most prevalent three-way interactions as indicated by the epistatic network.
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We used this pairwise epistasis model per metabolite because we had previous evidence that RIL populations have a significant false negative QTL detection issue and wanted to be inclusive of all possible significant loci (Chan et al., 2011).
None of the pairwise epistasis tests among the significant and suggestive QTL was significant after Bonferroni correction.
This uses the pairwise maximum likelihood estimation procedure.
For metabolites epistatically affected by M.I.83 × M.IV.23 × cytoplasmic three-way interaction, we plotted the center of phenotypes corresponding to interaction of nuclear loci M.I.83 × M.IV.23 as estimated using the three-way epistasis and pairwise epistasis models.
Using pairwise epistasis model run for separately for all metabolites, we identified all significant main effect markers and, epistatic interaction terms.
To test for cytonuclear epistatic interactions, we used a pairwise epistasis model that directly tested all pairwise combinations of the 14 nuclear QTL and cytoplasm against all metabolites while including all terms from the additive model.
Tests for pairwise SNP interactions were performed using the epistasis command in PLINK.
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