Autoclave to kill RNases/DNases? - argument (Dec/14/2004 )
Due to the numerous contrary opinions amongst people in my lab, I have decided to put this question on the forum: does autoclaving destroy DNases and RNases? Half the people I know say it does, half say it doesn't....does anyone know definitively? Thanks 
If it dose kill RNase, why we need to treat water with DEPC and then autoclave it?
Autoclaving is not effective at eliminating RNase in solution because the RNases simply renature as the solution cools.
Read this http://www.ambion.com/techlib/tb/tb_178.html
I agree with the RNase issue (though other stubborn individuals do not), but I was a little less sure regarding DNases....
Autoclaving is not effective at eliminating RNase in solution because the RNases simply renature as the solution cools.
Read this http://www.ambion.com/techlib/tb/tb_178.html
You should read your link, it says:
"Autoclaving inactivates enough of the RNase A to protect the probe from degradation up to a concentration of 1 µg/ml. Note that only a portion of the RNase is inactivated by autoclaving, otherwise the RNA probe would remain intact at any RNase concentration. Autoclaving alone may be sufficient to eliminate enough RNase for some applications."
To be on the safe side, just use DEPC in a concentration of 0.1% (NOT tris or hepes, or other reagents with amine groups). Otherwise be sure to autoclave your solutions several times.
DNases yes, RNases no
We used to autoclave our RNAse A to sterilise it. Didn't (seem to)lose any functional activity
Yes, so did I too, and it was still active.
however I never compared the activity of the autoclaved RNase versus the non autoclaved.
I cannot say that autoclave doesn't hurt RNase at all, but I can tell you autoclave is not enough to get rid of RNase.
i agree RNase could not be removed.