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To explore the full evolutionary dynamics of our model with non-antigenic and antigenic mutations, we implemented an individual-based model that allowed for an arbitrary number of new strains to enter the population and co-circulate.
To confirm whether or not the properties can be used as criteria for distinguishing LoF and GoF mutations, we implemented a classification technique using the support vector machine, random forest, and linear logistic regression methods with 50 data sets which contain equal numbers of LoF and GoF mutations to avoid bias.
To determine whether spindly HA phylogenies, co-circulation of antigenic variants, and high annual attack rates indeed come out of a model that incorporates deleterious mutations, we implemented a phylodynamic model that simulates the occurrence of both non-antigenic (largely deleterious) mutations and antigenic mutations ('Materials and methods').
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For synonymous mutations, we have implemented the Codon Adaptation Index.
Though there were constraints on the kind of "mutations" we could implement, we were really required to think hard about the consequences of each action, rather than just mindlessly follow a protocol and answer some questions thereafter.
In order to test the effect of the mutations on protein folding, we implemented the GFP-folding assay.
To provide a systemic gateway for mutation-disease relationship exploration, we implemented several search features on the website, including mutation description syntaxes (e.g. 'p.Gly1349Asp'), reported gene name (e.g. 'CFTR'), chromosomal region (e.g. 'chr1 100000-200000'), disease (string search or multiple option search), or OMIM disease IDs.
For example, we implemented oriented mutation operators for RNA secondary structure by specifically targeting mutations to bases that are predicted to be paired or unpaired, to, respectively, decrease or increase the strength of the mutated RNA structure.
To extract read count from a sequencing platform where the mutation was not detected automatically, we implemented a "matrix" code which counts reads not flagged as optical/PCR duplicates and have base quality ≥ 15 at the queried variant site.
To identify pathways when simultaneously integrating gene expression, CNV, somatic mutation data, and methylation data, we implemented PARADIGM [ 15], an algorithm that predicts individual tumor pathway activity by factor graph.
We implemented such in silico mutation by changing parameter inequalities for XylRa-dependent XylS expression and for the capability of high levels of XylS (i.e. XylSh) to activate Pm (see Table 1 and Methods section).
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