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The analysis of ChIP-seq samples outputs a number of enriched regions (commonly known as "peaks"), each indicating a protein-DNA interaction or a specific chromatin modification. When replicate samples are analyzed, overlapping peaks are expected. This repeated evidence can therefore be used to locally lower the minimum significance required to accept a peak. Here, we propose a method for joint analysis of weak peaks. Given a set of peaks from (biological or technical) replicates, the method combines the p-values of overlapping enriched regions, and allows the "rescue" of weak peaks occurring in more than one replicate.

MSPC comparatively evaluates ChIP-seq peaks and combines the statistical significance of repeated evidences, with the aim of lowering the minimum significance required to “rescue” weak peaks; hence reducing false-negatives.

MSPC can be used from:

About#

With most peak callers (e.g, MACS), the false-positive rate is a function of a user-defined p-value threshold, where the more conservative thresholds result in lower false-positive rates---the penalty of which is the increase in the number of false-negatives. While several probabilistic methods are developed to jointly model binding affinities across replicated samples to identify combinatorial enrichment patterns (e.g., jMOSAiCS, or this, or this, or this), the more commonly used peak callers (e.g., MACS) operate on single samples. Additionally, the choice of peak caller is generally an established step in a ChIP-seq analysis pipeline, since altering it may require changes to the data pre- and post-processing of the pipeline. Therefore, the remainder of this manuscript is focused on the post peak calling methods to lower false-positive rates as they can be applied on the output of any peak caller with minimal changes to the established analysis pipeline, and can combine evidence across replicated samples.

Motivation#

Ultimately, with most peak callers (e.g, MACS), the false-positive rate is a function of a user-defined p-value threshold, where the more conservative thresholds result in lower false-positive rates---the penalty of which is the increase in the number of false-negatives. While several probabilistic methods are developed to jointly model binding affinities across replicated samples to identify combinatorial enrichment patterns (e.g., REF1, REF2, REF3, REF4, REF5), the more commonly used peak callers (e.g., MACS) operate on single samples. Additionally, the choice of peak caller is generally an established step in a ChIP-seq analysis pipeline, since altering it may require changes to the data pre- and post-processing of the pipeline.