DNA-Prep Reagents Kit, RUO

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- cells were seeded at 5×104 cells/ml and treated with IC50 values of AFha, AFe and Cha and incubated for 16 and 32 h	- 100 µl lysis reagent were added to each tube and incubated for 5 min at room temperature.

Upstream tips
- cells were seeded at 5×104 cells/ml and treated with IC50 values of AFha, AFe and Cha and incubated for 16 and 32 h
Protocol tips
- 100 µl lysis reagent were added to each tube and incubated for 5 min at room temperature.

Publication protocol

HeLa cells were seeded at 5×104 cells/ml in 10 ml aliquots in 10 cm culture dishes and treated with IC50 values of AFha, AFe and Cha. The cells were incubated for 16 and 32 h. After the appropriate incubation period, the HeLa cells were trypsinized for 10 min, re-suspended in phosphate-buffered saline (PBS) and transferred to polypropylene tubes. The Coulter DNA Prep reagents kit was used for DNA cell cycle analysis, as per the manufacturer's instructions. Briefly, 100 µl lysis reagent were added to each tube and incubated for 5 min at room temperature. Thereafter, 500 µl PI (50 µg/ml) were added and the tubes were incubated for 15 min at 37°C. Flow cytometric analysis was performed directly following incubation. A Beckman Coulter Cytomics FC500 was used for all flow cytometricanalysis. FlowJo_V10 was used for analysis.

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