Yeast Cell DNA Content Analysis

Samples were prepared for flow cytometry, as described previously [29 (link)], with modifications according to specific strain requirements. Yeast cells were stained using propidium iodide (PI) (16 μg/mL) (Sigma Aldrich, St. Louis, MO, USA) (strains Y1023, Y1024) or SYTOX Green (0.5 µM) (Invitrogen, Carlsbad, CA, USA) (strains Y1006, Y1012, Y1013, Y1014, Y1017, Y1018, Y1019, Y1020). The DNA content was determined by measuring the PI or SYTOX Green fluorescence signal (FL2 or FL1, respectively) using Becton Dickinson FACS Calibur and CellQuest software (BD Bioscience, San Jose, CA, United States).

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Denkiewicz-Kruk M., Jedrychowska M., Endo S., Araki H., Jonczyk P., Dmowski M, & Fijalkowska I.J. (2020). Recombination and Pol ζ Rescue Defective DNA Replication upon Impaired CMG Helicase—Pol ε Interaction. International Journal of Molecular Sciences, 21(24), 9484.

Publication 2020

propidium iodide SYTOX Green FACS Calibur CellQuest software

Cells Flow cytometry Fluorescence Propidium iodide Strains Sytox green Yeast

Corresponding Organization : Institute of Biochemistry and Biophysics, Polish Academy of Sciences

Other organizations : National Institute of Genetics

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Variable analysis

independent variables

Staining method (propidium iodide or SYTOX Green)

dependent variables

DNA content, measured by PI or SYTOX Green fluorescence signal (FL2 or FL1, respectively)

control variables

Specific strain requirements (strains Y1023, Y1024, Y1006, Y1012, Y1013, Y1014, Y1017, Y1018, Y1019, Y1020)
Sample preparation, as described previously [29]

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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