GSM9595236: Y. lipolytica strain E122-EdU labeling without hydroxyurea for 45 minutes #1; Yarrowia lipolytica; OTHER
To identify early- and late-firing origins, stationary phase cells were transferred into fresh YPD containing either EdU+HU or EdU alone. A mock (no EdU) control was included. At each time point, replication was halted with sodium azide (0.1%), and cells were collected for flow cytometry and DNA extraction. DNA was purified using a Qiagen Genomic-tip after Zymolyase-20T (Sunrise Science Products, N0766391) digestion and lysis. Purified genomic DNA was fragmented using the Bioruptor Pico and checked on a Bioanalyzer (100–550 bp fragments). Biotinylated DNA was ligated to adapters and PCR-amplified using the Illumina TruSeq kit. The resulting libraries were sequenced to profile replication timing at high resolution.