DT40 Cell Synchronisation - (Jan/03/2012 )

Hey

I have recently been given DT40 cells to do some experiments with, but I need to synchronise these cells in S-phase. Everywhere I look recommends elutriation but I was hoping to do this using thymidine. Does anyone know if this works and if so a concentration of thymidine that would do so?

Any help would be greatly appreciated.

Debbie

It does seem to work for some cells, and a quick google search shows that about 2 mM is the concentration to aim for, but this will vary according to the cells you are using and whether you want all the cells to be in S or only a proportion (70% ish)?.

2.5 mM is what I use for U2OS osteosarcoma cells, so I guess 2 mM makes sense. It doesn't matter if all the cells are in S, its more to try and enrich for cells in S phase. 70 % would be great to get!
Thank you! :-)