could i use sterile deionized water instead of DEPEC water in RNA extraction - (Jun/09/2011 )
i am doing RNA extraction for doing RT-PCR but i dont have DEPEC in my lab.
so could i use autoclaved deionized water instead of DEPEC treated water
Short answer: No.
Long answer: it may be OK, but if you have any RNAse contamination in your deionised water supply, your RNA will be digested. Autoclaving will not prevent this - RNAses will renature.
You can buy RNAse free water if required.
you don't need DEPC, just order this water: Invitrogen Cat.No. 10977035, 12.6 EUR for 500ml
But i have another question
what about the micrfuge tubes and pipette tips (plasticwares)should i wash them by DEPEC water before autoclaving???
if you didn't order them "rnase free" then you can soak them in water with depc overnight, drain and autoclave (after soaking, you should also autoclave the water to eliminate the depc).
mdfenko on Fri Jun 10 18:41:56 2011 said:
(after soaking, you should also autoclave the water to eliminate the depc).
which water do u mean??????????????
Basically Mdfenko is saying that you can soak the tips etc in water with some fresh DEPC over night. You should then drain off the water and keep it and then autoclave both the water and the tips to get rid of any residual DEPC (which is extremely toxic to you and the environment).