BackgroundProtein-DNA binding sites of ChIP-seq experiments are identified where the binding affinity is significant based on a given threshold. The choice of the threshold is a trade-off between conservative region identification and discarding weak, but true binding sites.ResultsWe rescue weak binding sites using MSPC, which efficiently exploits replicates to lower the threshold required to identify a site while keeping a low false-positive rate, and we compare it to IDR, a widely used post-processing method for identifying highly reproducible peaks across replicates. We observe several master transcription regulators (e.g., SP1 and GATA3) and HDAC2-GATA1 regulatory networks on rescued regions in K562 cell line.ConclusionsWe argue the biological relevance of weak binding sites and the information they add whenrescuedby MSPC. An implementation of the proposed extended MSPC methodology and the scripts to reproduce the performed analysis are freely available athttps://genometric.github.io/MSPC/; MSPC is distributed as a command-line application and an R package available from Bioconductor (https://doi.org/doi:10.18129/B9.bioc.rmspc).Supplementary InformationThe online version contains supplementary material available at 10.1186/s12859-023-05340-x.