Protocol Online logo
Top : New Forum Archives (2009-): : PCR, RT-PCR and Real-Time PCR

DNase treatment after RNA extraction using RNeasy kit of qiagen - DNaes treatment after RNA extraction (Jun/07/2011 )

Hello everyone,

I isolated plant leaf RNA using RNeasy kit of qiagen. Kit itself assure DNA contamination free RNA but some minut quantity I recorded on my agarose gel. I've not done on-column DNase treatment but now I want to treat my samples with DNase. Can I use NEB DNase for this RNA. second RNA cleanup procedure is required after this treatment or not? How much RNA I should treat with DNase?



Yes you can treat your isolated RNA with Rnase free DNase. You also need buffer to stop the reaction. How much RNA to treat with certain amount of DNase is important. You should read the instructions coming with the product.


We had also some DNA contamination after on-column DNase treatment with Qiagen kit. So we rather use Turbo DNA-free from Ambion, and treat the whole volume of RNA and recomended protocol. There is no cleanup procedure, you just centrifuge it to spin down particles containing the enzyme.