Hi every body
I added DEPC to water and then shake it vigorously and I saw that it mad foam. After autoclaving I did not see any foam. Is it natural?
Best regards
Very basic chemistry and it does not require any guessing. ;)
The "foam" you see are bubbles of CO2. It is perfectly normal and you should anticipate it; in fact, if you do not see the bubbles, you need to acquire a fresh batch of the reagent.
Diethyl pyrocarbonate (DEPC) is highly reactive, it modifies amino group containing AAs, hence its use in inactivating RNase, which is a very stable enzyme. DEPC breaks down into CO2 and ethanol in the presence of water. Ethanol evaporates when you autoclave the water.
It goes without saying that never DEPC treat any buffer or solution that contains NH2-groups, such as Tris. Use DEPC treated water to make the buffer with appropriate precautions to ensure that resulting buffer remains uncontaminated with RNase.
I imagine it's probably fine. I don't usually shake DEPC treated water. We use a stir bar + magnetic plate. You can see clear blobs of DEPC when doing this, which might be causing your foam. How long are you letting the DEPC water incubate before autoclaving?
Dear Samantha Jeschonek
I Shake it for about 5 minutes because of my curiosity. I incubate it for 12h at 37C in a shaker incubator (200 rpm) then I autoclave it two times (2*15min).
Very basic chemistry and it does not require any guessing. ;)
The "foam" you see are bubbles of CO2. It is perfectly normal and you should anticipate it; in fact, if you do not see the bubbles, you need to acquire a fresh batch of the reagent.
Diethyl pyrocarbonate (DEPC) is highly reactive, it modifies amino group containing AAs, hence its use in inactivating RNase, which is a very stable enzyme. DEPC breaks down into CO2 and ethanol in the presence of water. Ethanol evaporates when you autoclave the water.
It goes without saying that never DEPC treat any buffer or solution that contains NH2-groups, such as Tris. Use DEPC treated water to make the buffer with appropriate precautions to ensure that resulting buffer remains uncontaminated with RNase.
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