2 replies to this topic

[yekaterina poloz]

I use DEPC treated H2O for all of the buffers from electrophoresis to immunodetection during northern blotting. If I stop autoclabing DEPC treated H2O or buffers made from it, will it have a negative impact on the blots? I know carboxymethylation of purines by DEPC inhibits translation but what about migration in the gel, RNA:RNA or DNA:RNA hybridization or immunodetection? It's not that I am lazy but we've all of the sudden have a very limited access to an autoclave.

[bob1]

DEPC will break down in the presence of water and heat, you can just heat the solution for a while (I don't know how long would be suitable).  The major issue is of course that DEPC is quite toxic, so I am not sure that you would want it being realeased into the lab space every time you open the bottles for you to breathe in.

[btc8]

yekaterina poloz, on 23 August 2010 - 12:55 PM, said:

I use DEPC treated H2O for all of the buffers from electrophoresis to immunodetection during northern blotting. If I stop autoclabing DEPC treated H2O or buffers made from it, will it have a negative impact on the blots? I know carboxymethylation of purines by DEPC inhibits translation but what about migration in the gel, RNA:RNA or DNA:RNA hybridization or immunodetection? It's not that I am lazy but we've all of the sudden have a very limited access to an autoclave.

You need to autoclave it to inactivate the trace DEPC.  DEPC will react with anything that has a -N, -O, or -S, so it may screw up your RNA bases.  I would autoclave to be safe, especially if you're using it to store RNA in.

-b