title: GSM6674356: WT yeast, MnCl2 treatment, replicate 1; Saccharomyces cerevisiae; OTHER
library_construction_protocol: Extraction methods were based on previously published methods from our lab (data available at GSE142457, GSE166983, and GSE175553). In brief, cells were harvested by filtration, flash frozen, and lysed under cryogenic conditions. Samples were either left untreated or treated as described in the "treatment protocol" section. Pol I was immunoprecipitated as previously described. Finally, a total RNA extraction was performed using acidic phenol and chloroform. DNA libraries were constructed exactly as previously published (Jacobs and Huffines et al., "The small-molecule BMH-21 directly inhibits transcription elongation and DNA occupancy of RNA polymerase I in vivo and in vitro", JBC, 2021). Briefly, a DNA linker containing a UMI sequence was ligated onto the 3' end of RNA reads. Samples were precipitated, reads were reverse transcribed, and DNA was circularized. Circular DNAs were amplified, libraries were purified with PCRClean DX beads, and sequencing was performed.