GSM8641586: cdc73△ yeast, replicate 2; Saccharomyces cerevisiae; OTHER
Extraction methods were exactly the same as described in previous publications from our lab. To summarize, yeast cells were rapidly harvested via filtration and lysed under cryogenic conditions. Pol I was isolated by immunoprecipitation. Finally, a total RNA extraction was performed with an acidic phenol/chloroform extraction. NET-seq libraries were constructed exactly as described in previous publications by our lab. In summary, after the phenol/chloroform extraction, a DNA linker containing a UMI sequence was ligated onto the 3' end of resulting RNAs. Next, cDNAs were synthesized via reverse transcription. Finally, cDNA was circularized and amplified. Resulting libraries were purified with PCRClean DX beads and reads were sequenced.