rt-pcr

Jose de las Heras jose.delasheras at virgin.net
Mon Feb 14 15:32:39 EST 2005


"as" <nospam at spam.dk> wrote in message 
news:WB5Qd.128$Qx7.110 at news.get2net.dk...
> Hi
> I have a basic question regarding RT-PCR.
>
> I use Trizol for RNA purifacation, DNAse1 from SIGMA for removal of DNA
> contamination (15 minutes at 38 degree) and Superscript 2 for RT.
> At the last step I add RNAse inhibitor (together with the SS2),
> would it be possible to add the RNAse inhibitor at the DNAse1 treatment 
> step
> (i.e when disolving my RNA pellet in DEPC water), or will it also inhibit
> DNAse1 function?
> Thanks
> A.Salanti

Go for it. It'll be just fine. In fact, I had some problems with RNA getting 
degraded after DNAseI treatment, and adding a little of RNAse inhibitors 
allowed me to get good RNA after treatment. Now, as a matter of routine, 
when I have small samples of "precious" RNA I use inhibitors at every step 
to avoid heartbreaks.

Jose 





More information about the Methods mailing list