Nuclease free water means free from both nuclease. DNase is normally not very stable. It can be deactivated by vortexing or heating but RNase are more stable. In case of RT you either use RNase free water or nuclease free water.

Hi Ubanaco,

As mentioned by Ganapati, nuclease free water is a water free of RNase and DNase.To obtain nuclease-free water you will need to treat your water with 0.1% DEPC (diethyl pyrocarbonate-Sigma, cat # D5758, stored @ 4˚C).

Here it is the protocol: Add 1mL DEPC in 1000 mL Milli-Q water. Stir overnight at 37˚C and autoclave next day to evaporate DEPC. If you don't want to go through this work, you can just by DEPC-treated water from different companies. Here it is the catalogue number from Ambion (#9906).

Good luck in your experiments.

Thank you so much Viviane

Nucelase = DNAase and RNAase., so we can use this Nuclease free water for both RNA work and DNA Work. Especially in NGS Library preparation.