time of DNase I treatment - (Sep/08/2008 )

Hallo to everyone!

I would like some suggestions about the incubation of RNA with DNase I enzyme for cleaning!
I use the enzyme from New England Biolabs and they propose 10 min treatment, but for me it didn't proved sufficient...
Any better idea?

Thanks
ilianaki

We usually do 20 min at 37C.

Hallo to everyone!

I would like some suggestions about the incubation of RNA with DNase I enzyme for cleaning!
I use the enzyme from New England Biolabs and they propose 10 min treatment, but for me it didn't proved sufficient...
Any better idea?

Thanks
ilianaki

We use ambion's dnase and the incubation is 40 min.

Thank you both!
I will try!

Incubation time depends on enzyme amount used and the amount of DNA to digest. If you have a high quality enzyme 10-20mins usually are enough when using 1-10U of enzyme.
It is important to have an incubation buffer with the necessary cofactors (CaCl2, MgCl2)!
You should definetly use a DNase which is really RNase free (and optionally add an RNase Inhibitor like the Protector RNase Inhibitor).
Good success!!!