Optimizing Trizol mRNA extraction - (Jan/24/2012 )

Dear forum members,

I was wondering if you could contribute some ideas on how to optimize Trizol mRNA extraction and minimising as much genomic DNA contamination as possible? Any other methods apart from DNase digestion would be much appreciated.

Cheers!

-Byong-

No RNA prep method perfectly excludes DNA contamination however, discoverer of Trizol have launched a new reagent formulation which is claimed to avoid any DNA contamination.

I have not tested it yet

http://www.mrcgene.com/rnazol.htm

http://www.lifetechindia.com/RNAzol_RT_Protocol_Life_Technologies_India.pdf

http://www.nature.com/nmeth/journal/v7/n12/full/nmeth.f.315.html

-Kamran-

Hello~ Thanks for replying! It looks interesting but I'm afraid I won't have a chance to use it. I think my supervisor would rather I stick to Trizol as opposed to ordering new materials in . :9

-Byong-

There is a an alternate step spinning the Trizol before adding chloroform, that should spin down the high-weight DNA and solid particles. You can test if that helps.
Also we take only upper 2/3 od aqueous phase to limit DNA contamination from the interphase.
Depends what application you want your DNA-free RNA for. If it's PCR, just give up and buy a DNase.

-Trof-