Each spot assay was performed as follows: each mutant was grown in 4 ml of YPD liquid media until OD600 reached ~2.0. Cell density of the yeast culture was normalized to OD600 of 0.2 to ensure equal plating. Four five-fold serial dilutions were then performed for each strain in a sterile 96-well plate using sterile deionized water as diluent. 3 μl of each dilution was spotted onto either YPD or YPD plates containing either hydroxyurea (HU) or methyl methanesulfonate (MMS) of indicated concentrations. All three plates were incubated at 30°C for 2 to 3 days before the representative images were acquired using a Bio-Rad ChemiDoc MP imaging system.

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