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These are the most frequent base substitutions induced by reactive oxygen species, byproducts of normal aerobic metabolism generated at high levels in all inflammatory processes and after exposure to a wide variety of carcinogens and toxicants [27], [28], [29], [30], [31], [32], [33], [34].
The most frequent base was taken as the consensus for the other polymorphic sites.
Subsequently, variance of the most frequent base in that position is calculated from the base composition.
The most frequent base at a position was chosen for the consensus sequence.
If there were more than two overlapping fragments the most frequent base was used to determine the consensus.
The most frequent base changes were C → T and G → A, each in 17 cases, mainly at hotspot sites.
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A polymorphism was predicted when the second model that allowed the population to be an arbitrary mixture of the two most frequent bases at this position explained the data better by a likelihood ratio test (E-value ≤0.01).
Fittingly, the most frequent base-determinator is A (the second most frequent being G), and its complement is U (note that U can form complementary pairs with G as well).
The most frequent base-substitution was G C to A T, 141 for the entire genome most of which were on chromosomes I or X, 55 and 31 respectively.
Polymorphisms were predicted by applying the model implemented in the breseq pipeline that allowed the population to be, in each position, an arbitrary mixture of the two most frequent bases.
As we were interested in identifying polymorphic sites (PSs) where the mutations could occur in only a fraction of H. bizzozeronii population, we applied a model using the Breseq computational pipeline (Barrick et al. 2009), which allowed the population to have an arbitrary mixture of the two most frequent bases at each position in the genome (Jerome et al. 2011).
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