how to make nuclease free water ?

Chris LaRosa clarosa at biocomp.unl.edu
Tue Jul 11 10:58:30 EST 2000



Axel Schwekendiek wrote:
> 
> Dear all,
> 
> could anybody provide me with a short protocol on how to prepare nuclease
> free water? I need it for PCR, RT-PCR and RNA extraction from plant tissues.
> We have a milli-Q system and I don't want to use DEPC.
> 

If you have a properly working milli-Q system,,, simply add water to
sterile dispo plastic tubes etc.  The milli-q system puts out nuclease
free water. Be sure not to use reused washed glassware as the rnases
cannot be removed easily.  Be sure to dissolve final rna preps in
formamide as opposed to water.   This greatly reduces any residual rnase
activity.   Dont worry about inactivating rnases during the extraction
steps,  the plants release plenty which should be delt with by the
denaturing agents in the extraction buffers (phenol ,, guanadidium
depending on the particular procedure.






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