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The prevalences and their 95% confidence intervals (CI) were calculated by the exact binomial method and the differences between proportions were analysed using the Chi2 test.
To determine whether this increase is due to high false positives or due to the improved inference, we investigated the difference between callings provided by the binomial method and the Bis-class methods further by several different approaches.
For each model, we used NCSS statistical software (NCSS, Kaysville, UT, USA) to calculate the specificity and sensitivity required to construct ROC curves and estimate AUC (binomial method) and the 95% confidence interval (95% CI) for each of the three models.
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Both the log-binomial method and the Poisson method are generalized linear models with a log link function, which is assumed to be the correct form.
FDR-corrected q-values and posterior odds for each position of this locus are provided in the Additional file 5. Second, we did the following experiments to directly assess the difference between the binomial method and Bis-Class when the numbers of reads is reduced.
This value is just over three times as large as we obtained from the binomial method, and 6.5 times as large as from the Poisson method.
We demonstrate that Bis-Class alleviates the problems of the binomial method and improve sensitivity and accuracy using extensive simulations as well as analyses of actual methylC-seq data.
In the non-homogenous, clustered genomes, Bis-Class (solid green bar) outperforms the binomial method and exhibit much higher sensitivity (therefore lower false negatives) than the binomial method (solid purple bar, Figure 3).
We then used the binomial method and Bis-Class for methylation calling.
In genomes where DNA methylation occurs uniformly ('homogeneous'), both the binomial method and Bis-Class provide similar results across almost all settings (purple and green bars filled with diagonal lines in Figure 3).
† The intercept estimate was -1.5147 for the log-binomial method and -1.8311 for the Robust Poisson method.
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