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Most sequence variations observed in the NGS reads by indexed amplicon sequencing reflect mutations well in the library in comparison with the HRM analysis data.
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Leslie Brunetta and Catherine Craig also explain the mechanisms behind evolution (natural selection, gene flow, genetic drift, mutation) very well, in addition to colloquially mentioning all the pre- and post-zygotic barriers to reproduction through the world of spider speciation.
For proteins that have evolved by the sequential accumulation of point mutations, most of those mutations must work well in many different contexts.
Specifically, polony technology was successfully applied to both detect intragenic mutations in well-defined mutational hotspots in key cancer genes as well as determine if loss of heterozygosity of these same genes had occurred.
This may contribute to a bias with greater attribution of disease causation to mutations in well-represented and well-covered genes.
In a proof of concept study, we have rediscovered known mutations in well characterized cancer cell lines.
Fifteen years later, mutations in well over 50% of the 275 amino acids of subtilisin have been reported in the scientific literature.
However, it seems unlikely that so many different missense mutations as well as in-frame deletions could cause the same gain-of-function effect.
However, each of these datasets has its own biases; in particular, they are enriched for mutations in well-known genes that have been widely studied in cancer.
Seven were intergenic or within an intron, the remaining were missense/nonsense or frameshift mutations in well-characterized genes that are not associated with mutator phenotypes.
In the renal carcinoma tissue harbouring the A3243G mutation as well as in the two other renal carcinoma tissues investigated, lack of VHL protein was observed.
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CEO of Professional Science Editing for Scientists @ prosciediting.com