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Autoclave reagents for rna? - (Jun/04/2007 )

Hi there,

I am planning to concentrate an rna sample using a Na Ac protocol. I made up the 3M Na Acetate and the glycogen co-precipitant in depc treated h2o in sterile tubes.

Is it also necessary to autoclave them ?

thanks

-smurray-

for sure i would autoclave.

-fred_33-

it doesnt matter whether you keep your reagent in a sterile tube......you must autoclave your reagent first.

-T. reesei-

you need to DEPC treat the solution, not just the water you use to make it. This is true for all slns that do not contain Tris (because DEPC interacts with Tris (i think it is amines) you cannot DEPC treat these slns so you do the best you can by using DEPC treated water) of course, after DEPC treatment you always autoclave to seperate the DEPC into carbon and water so it does not affect your reactions etc. Anyway, make it up with regular water and DEPC treat the whole solution.

-beccaf22-

Note that autoclaving won't destroy RNAse. Only DEPC or baking overnight can do that. So the main point of autoclaving after adding DEPC is to destroy the DEPC.

-Zouden-

QUOTE
make it up with regular water and DEPC treat the whole solution.


Hey thanks. What concentration of DEPC do you use?

cheers

-smurray-

QUOTE (smurray @ Jun 6 2007, 04:48 AM)
QUOTE
make it up with regular water and DEPC treat the whole solution.


Hey thanks. What concentration of DEPC do you use?

cheers



To prepare RNase free water or other aqueus solution Add DEPC to 0,01% (v/v)

cheers Alice

-sisma-