rt-pcr

as nospam at nospam.com
Sun Feb 20 08:14:59 EST 2005


Ok thanks, I will try
"Jose de las Heras" <jose.delasheras at virgin.net> wrote in message 
news:37cgj0F5a45a9U1 at individual.net...
>
> "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
> 





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