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The conservation frequency differed a great deal among pair types in maize, where divergent pairs are conserved at rates close to half that of convergent or tandem pairs.
Gene and domain effects were estimated separately for LQTS and BrS, but the effects of SIFT, PolyPhen, Grantham, conservation, frequency and inframe variants were estimated jointly across the outcomes.
The hexamers selected were such that they have a conservation frequency in mouse similar to that found for the set of ESEs in our exon set (see Methods for details).
From the obtained hexamers, we selected 10 times two independent random sets of 238 and 380 hexamers, respectively, with a conservation frequency similar to that of ESEs [see Additional File 1].
We then predicted the (dis)assembly pathway for each randomly reweighted complex and assessed the conservation frequency, and repeated this process many times in order to calculate the intrinsic probability of assembly conservation.
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In every comparison, divergent gene pairs were conserved least often, with conservation frequencies significantly below the all pair average.
We therefore extracted hexamers and their conservation frequencies from the CDS of single-exon genes conserved between human and mouse.
The three classifications (i.e., prediction tools, conservation, and frequency) were combined according to the following rules: If the prediction based on frequencies was "benign," then the final classification of the variant was "probably benign" regardless of the other predictions.
In brief, classification of pathogenicity was performed using a combination of prediction programs, evolutionary conservation, and frequency data.
The functional implications of the conservation or frequency of SSRs, if there are any, also unfortunately must remain unclear at this stage.
These differences could reflect selection on recombination of alleles linked to uptake sequences, but they might also reflect different levels of conservation and/or frequencies of horizontal transfer in these groups.
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