Does anyone elute RNA in 1mM EDTA? Downstream protocol has a 70ºC RNA denaturation step and the manual suggests to use EDTA to avoid RNA degradation?
I am going to perform an RNA clean up using RNeasy spin columns, and I would like to elute RNA in 1 mM EDTA instead in DNase RNase free water, but I am not sure if that it is possible. Otherwise, has anyone denatured RNA in water at 70ºC without experiencing RNA degradation? Thanks a lot!
I elute my RNA in TE-like buffer that includes 10mM TRIS pH7.5 and 0.1mM EDTA. EDTA chelates metal ions (Mg2+) that otherwise are required for exonuclease activity (so that will prevent degradation of your RNA). However if you use too high concentrations of EDTA you might have issues with downstream applications, e.g. RT-PCR (RT enzyme requires Mg2+ to work). My elution conditions prevent the degradation and at the same time you should not have any issues with downstream analyses.
The best is to use deionised water treated with DEPC to avoid RNA degradation.
Regards
I elute my RNA in TE-like buffer that includes 10mM TRIS pH7.5 and 0.1mM EDTA. EDTA chelates metal ions (Mg2+) that otherwise are required for exonuclease activity (so that will prevent degradation of your RNA). However if you use too high concentrations of EDTA you might have issues with downstream applications, e.g. RT-PCR (RT enzyme requires Mg2+ to work). My elution conditions prevent the degradation and at the same time you should not have any issues with downstream analyses.
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