During RNA extraction (using RNeasy kit), is an on-column Dnase treatment useful when you also performe DNAse treatment (Invitrogen) afterwards?

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What is the best concentration of cDNA to be used for qPCR?

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Michael Porter

DNase treatment is usually quite thorough, I've never seen it go wrong. I wouldn't have even thought about doing it twice.

Imran Sabri

DNase treatment is enough. you need not to do both.

What you can do is be perfect in rather than wasting time in the second one.

Vincent Oustric

One Dnase treatment is enough. I usually perfome this after the extraction just before the retro transcription.

Jörg Vogt

I fully agree with Vincent Oustric's answer.

btw: for solving RNA/DNA after any procedure, I used ddH2O or DEPC-H2O .. So I have no doubt that downstream applications might be inhibited by the solution buffer's ingredients