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DNase I treatment of RNA dissolved in TE buffer - (Aug/04/2005 )


I want to treat my isolated RNA with DNase I , but the RNA was suspended in the 40μl of TE buffer pH 8.0, so I dont know whether the EDTA will exert bad influences to the DNase I
Thanks for your help in advance.

weimin

-weimin42-

Hi,

DNase requires Magnesium ion (~3mM) for its activity. EDTA, being a chelator, presence will surely not allow complete DNase activation.
Why not u reprecipitate ur RNA and suspend in Sterile water and perform DNase I digestion.


Nidhi

-nidhi-

QUOTE (nidhi @ Aug 4 2005, 10:43 AM)
Hi,

DNase requires Magnesium ion (~3mM) for its activity. EDTA, being a chelator, presence will surely not allow complete DNase activation.
Why not u reprecipitate ur RNA and suspend in Sterile water and perform DNase I digestion.


Nidhi


OR as the EDTA in TE is only 1mM you could just add an extra 1mM Mg to the mix to counteract the EDTA

-John Buckels-