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Preserving RNA - How to store lised cell (Aug/03/2005 )

Here in my lab we store cells lised in " RLT" buffer from Qiagen. We can't :( store them at -80ºC, so we add 2-mercaptoethanol, wich I personaly HATE! :wacko:
Someone, please, give me an alternative!!! :(


Look a few post down for a thread named "Homemade RNAlater".

I hate beta-mecaptoethanol too. Try using DTT -much less smell smile.gif


DNA sequencing software

-Daniel Tillett-

For storing lysed cells to later retrieve RNA I use Tri-Reagent or Trizol, you just lyse the cells in tri... and place at -80C directly RNA is stable for as long as I have tried to store...6 months plus... I like doing the RNA isolation this way too, have always gotten better (ie: better yield, not degraded) RNA from this method than with any kits...

BTW I personally don't like RNAlater... Too much salts, you shouldn't lyse your cells in RNAlater because it works best if you can pellet whole cells out and remove the RNAlater before doing your extraction (again b/c of all the salts)



alternatively, if you don't have tri, you can store your cells washed by PBS at -80. And before lysing, you heat at 50°max your lysis solution and apply directly on frozen pellets. Resuspend carefully and quite completely with a p1000 (or less if applicable) but p1000 is better than all pipettes even if it's a 1ml pipette.
Frozen pellet can be stored for weeks at -80° without degradation, if the pelleting procedure is relatively quick. (i mean detach cells or collecting cells and a wash with PBS). I've never tried a quick frozen of pellet in nitrogen,but i'm gonna doing this in my next exp.


The post metionned by Daniel is available here