GSM5861250: rpa190-F1205H 2; Saccharomyces cerevisiae; OTHER
Extraction methods were based on previously published works from our lab (data available at GSE142457, GSE166983, and GSE175553). Briefly, cells were harvested via filtration, flash frozen, and lysed under cryogenic conditions. Immunoprecipitation for Pol I was performed as previously described and RNA was isolated via acidic phenol/chloroform extraction. DNA libraries were constructed 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). In brief, a DNA linker including a UMI was ligated onto the 3' end of RNA reads. Samples were precipitated, and reads were reverse transcribed. Finally, DNA was circularized, amplified, and sequenced.