RNase h electrophoresis - (Jun/10/2012 )

All,

Am new to this, but have utilized the site often for protocol research.

I am looking to remove RNase h from a sample and am unsure if electrophoresis will do the trick, or if there might be a better method.

I am not sure if simple de-naturation via high temp in a PCR would also help.

I am planning on utilizing native PAGE gel, probably 10% to start, with post run staining (Sybr AU) as our lab is not to keen on using EtBr.

We are hoping to synthesize a long ss DNA scaffold for some further research projects.

Hope I have provided enough initial info here.

Let me know if there is more data that needs to be provided.

Ciao

Donald K

donaldkellis on Sun Jun 10 15:17:48 2012 said:

Welcome

for what purpose? If it is to just clean up DNA, a simple phenol-chloroform extraction or a DNA kit extraction will work fine.

No - RNases are very good at re-folding post denaturation.

That process should work, but you also need to think about how you will elute the DNA from the gel. It would also pay to have a close look at the % gels for DNA and maximal resolution. I don't think that DNA gels will be denaturing enough to stop RNases.