The effect of metal cations on the capture of Pol II-derived nascent transcripts in NET-seq experiments

The goal of this study was to test the effect of metal cations (CaCl2 and MnCl2) during Pol II NET-seq. These metals are readily used in the majority of studies that use NET-seq to analyze Pol II occupancy, but their effect on nascent transcript capture has not been analyzed. Our results suggest that the inclusion of these metals in Pol II NET-seq experiments does not cause a significant change in Pol II occupancy in the untreated control vs. metal-treated (CaCl2 or MnCl2) samples. Pol II NET-seq was performed in triplicate for each of three treatment conditions: untreated, CaCl2-treated, and MnCl2-treated.