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While some interactions compensate fitness losses, the effect on the relative mutant frequencies was small so that epistasis as a buffering mechanism for fitness losses might be rather inefficient.
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Identically, using the lacI mutation detection system, the mutant frequencies were 6.4±3.1×10−5 and 5.8±2.0×10−5.
This indicates that lacZ mutant frequencies are not correlated with a specific DNA repair defect and eventual tumor outcome, at least not in the experimental design presented here.
The study did not show any mutation or adduct induction in the exposed group compared to the control group: cII mutant frequencies were 11.0±4.5×10−5 and 11.0±4.8×10−5 in control and exposed lungs, respectively.
For example, without AZT when epistasis is highest, the relative mutant frequencies are around 10−8 to 10−9 and epistasis affect those frequencies up to 3-fold (Figure 4 and Table S3).
However, because fluctuation analyses are laborious and the present study comprised 238 P. aeruginosa isolates, we took advantage of the fact that mutation frequencies and mutant frequencies are proportional in sufficiently large cultures.
Mutant frequencies were determined as the number of colonies on the selective plate divided by the number of colonies on the titer plate (times the dilution factor of 1,000).
The lacZ mutant frequencies were then calculated by determining the ratio of mutant pfu to total pfu.
The mutant frequencies were used to calculate the number of mutation events and the mutation rate, using Equation 3 above.
Spi− mutant frequencies were calculated by the number of Spi− mutants divided by the number of plaques on XL1-Blue MRA.
With the chromosomal assay used here, the mutant frequencies were too low (∼10−9) to measure coincident double mutants, which occur ≥3 logs less frequently (Bull et al., 2000).
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