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Genes with FDR<0.05, fold difference >1.5 and mean log intensity >2.0 were considered to be significant.
The antigen reactivity was defined by the mean log intensity measures of at least 4 replicate spots for that antigen on each slide.
To apply this method, we first computed the mean log intensity and the standard deviation for the replicated spots within each channel.
A 45° counterclockwise rotation of the (log2Cy3, log2Cy5 -coordinate system and a scalog2Cy5 -coordinateatesystem andlied to plot the log intensity ratio M=log2(Cy3/Cy5) vscalingean lof inthesity A=log2√Rcoordinates
The RNA degradation plot displays the mean log intensity averaged over all probes with the same index k of one microarray as a function of the probe index, k = 1,…,Npset.
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These proteins are listed in Tables 1 and 2 along with their mean log-intensity ratios, P-values, and FDRs.
The resulting data set was summarised with outliers removed to obtain mean log-intensity and standard error for each probe/array combination.
M-A plots were constructed for each slide, where the log-intensity ratio M = log(Cy3/Cy5) = [logCy3 - logCy5] were plotted against the mean log-intensity A = [(logCy3 + logCy5)/2] as described by [ 53].
M-A plots were constructed for each slide, where the log-intensity ration M = log(Cy3/Cy5) [logCy3-logCy5] were plotted against the mean log-intensity A = [(logCy3+logCy5)/2] as described by [ 41].
Quantile normalisation was applied to mean log-intensities in order to make the distributions essentially the same across arrays.
The test compares the differences in mean log-intensities between classes relative to the expected variation in mean differences computed from the independent samples.
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