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Differences in medians for non-normally distributed continuous variables for each exposed group and urban background were calculated using Wilcoxon tests with a post-hoc Bonferroni correction for multiple comparisons.
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For ChIP analysis, fold enrichment of specific enrichment over background was calculated using values normalized to input.
For each PCR, the specificity of the amplifications was validated and the threshold cycle above background was calculated using Bio-Rad iCycler software.
The statistical significance of these data, across the different dm genetic backgrounds was calculated using the two-tailed z test (Appendix 1 in Additional file 3).
In brief, background-subtracted intensities were calculated using foreground and background median signals, and normalized with the quantile normalization method.
Relative hybridization intensities and background hybridization values were calculated using Agilent Feature Extraction Software (9.5.1.1) (Agilent Technologies).
For each spot, background-subtracted median intensities were calculated using the median local background, and the signal intensity at each spot was normalized by the corresponding relative amount of double-stranded DNA.
After background correction, glutamate concentrations were calculated using the glutamate standard curve.
Background-subtracted mean intensities were calculated using the minimum method and further normalized using within-array loess normalization.
Correction of background intensities of Cy3 and Cy5 were calculated using median-feature and median-local background intensities of the uploaded file.
Intensity and background data for each element were calculated using Imagene 5.5 (BioDiscovery Inc., Australia).
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