FxCycle™ PI/RNase Staining Solution

Protocol tips

Upstream tips	Protocol tips	Downstream tips
		- Do not wash the cells after staining with FxCycle PI/RNase Staining Solution

Downstream tips
- Do not wash the cells after staining with FxCycle PI/RNase Staining Solution

Publication protocol

For cell growth measurements, 30,000 parental HCT116 and RKO cells along with three ZBED6−/− clones per cell line were seeded in 12-well plates in 10% FBS or 1% FBS, respectively, and were cultured for 13 d with real-time measurement of cell density every 10–12 h using an IncuCyte instrument (Essen Bioscience). To assess clonogenicity, 1,000 cells per well of parental cells, three ZBED6−/− clones, and two ZBED6−/−- AcGFP-Zbed6 clones were seeded in triplicate in six-well plate wells and cultured in McCoy’s 5A medium with 10% FBS for 2 wk. The resulting colonies were stained in methylene blue and counted. Cell-cycle analyses were performed using FxCycle PI/RNase (LifeTechnologies), and cells were analyzed on an LSRII flow cytometer (BD).

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Manufacturer protocol

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