2 replies to this topic

[megashock]

Hi all,

I have cells frozen in FBS+10% DMSO.

Is it possible to extract and to purify RNA from these cells?
Is it necessary to freeze the cells in PBS intead of FBS+DMSO in order to extract RNA ?

I can use Qiagen RNeasy kit, or trisole.

Thanks in advance for the help.

[doxorubicin]

Your cells are in a commonly used cryopreservation solution. If they were frozen slowly (in styrofoam or in a cryocylinder), they should be intact and viable.  You should just be able to spin them down, and follow the RNeasy manual.

[philman]

10% DMSO in FCS is what is usually used to preserve cells for freezing. Personally I would want to thaw and grow them up first before extracting RNA from them, as I would assume the duel shock of being exposed to a toxin (DMSO) and freezing would affect RNA expression somewhat, but the cells should still be viable if they were frozen properly.