DNAse I treatment of RNA - (Dec/07/2005 )

hi everyone

I have isolated total RNA using TRI reagent and would like to do DNAse I treatment(Sigma), i would like to know whether to re-do RNA isolation steps following enzyme digestion OR simply to perform phenol -chloroform extraction, also please let me know how much of EDTA to be added.

Thanks in advance..
agiesh

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You only need to add enough EDTA to soak-up all the magnesium in the dnase reaction. Usually 20mM is more than enough.

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hi
Thanks for the information provided,
my reaction condition is 6.25 mM Mgcl2 and 10 U of Dnase
do i need to include any buffer which contains ca2+ ions?.....
thanks in advance

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