DNase treat my RNA prior to Generacer exp? - (Apr/23/2007 )
hi all, i am trying to pull out the full sequence of a gene using Generacer from invitrogen. my question is, do i have to DNase treat my RNA prior to doing the 5' and 3' reactions?
the reason i ask is i got two lovely bands using trizol extraction. then treated with DNase but when i ran this reaction on the gel it was a smear. WHY??
i carried on with the generacer protocol using non treated RNA, treated \rna and control RNA anyway. i have not finished the experiment yet so do not know if they have worked. can i trust the non treated RNA ??
I think it is pretty important to get rid of any DNA trace from your sample before doing any sensitive application.
Hmmm the smearing from your sample can mean few things,
1st: It can be caused by RNase activity being introduced during pipetting
2nd: The initial sample is DNA and not RNA. Smearing means the DNase works on the sample.
(Obviously I wish it is the the latter one)