Guava Cell Cycle Reagent for Flow Cytometry

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	- The fixed cells were centrifuged at 800 Xg for 5 min at 4 °C - Suspended cells were treated with RNase A at 37 °C for 1 h followed by staining with Reagent.

Protocol tips
- The fixed cells were centrifuged at 800 Xg for 5 min at 4 °C - Suspended cells were treated with RNase A at 37 °C for 1 h followed by staining with Reagent.

Publication protocol

Control and transfected cells were harvested by trypsin (0.25%). Cell pellets were washed with cold PBS and then added drop-wise to pre-chilled 70% ethanol. The fixed cells were centrifuged at 800 Xg for 5 min at 4 °C, washed with cold PBS twice and then re-suspended in 0.25 ml PBS. The cells suspensions were treated with RNase A at 37 °C for 1 h followed by brief centrifugation to discard supernatant. Cells were re-suspended in 200 μl of Cell Cycle Guava reagent (Millipore, Billerica, MA, USA), incubated for 30 min in dark, and analyzed by EasyCyte Guava cytometer (Millipore). The data were further analyzed using FlowJo software.

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