How to confirm DEPC is inactivated after autoclaving it? - (Oct/25/2006 )
I am introducing RNA work in a lab and unfortunately RNA is degraded, even the ladder which was new.
Now, I am wondering if RNA degradation can be due to active DEPC in MOPS buffer, and following the loading buffer?
When I washed some equipment with DEPC treated water which was autoclaved I noticed a strong DEPC smell. I cannot remember that DEPC treated water smelled that strong but I'm also not sure if I just didn't noticed before. I mean normally DEPC should be degraded after autoclaving into EtOH and CO2 making water acidic. I measured pH but it was basic.
Unfortunately, this was my first time I prepared DEPC treated water and I'm not sure if the smell is normal or indicating active DEPC. Here is my treatment:
0.1% DEPC in water for 1h at 37°C or at RT over night. After Treatment Water was autoclaved.
I didn't do the autoclaving by myself but tape changed color, so it should be ok. Pleas help me
How to confirm DEPC is inactive and can active DEPC degrade RNA?
I prepare DEPC water by stirrring overnight. Then I was told to autoclave TWICE to ensure that all the DEPC is removed. I've never smelled it, but if you prepare this way all the DEPC should be inactivated.
usually i prepare DEPC water at 37°C over night, and then autoclave once. it works
it seems that active DEPC will not degrade RNA. I have used a bottle of active DEPC water(I thought it had beed autoclaved) to dissolve RNA, and the RNA was not degraded.