RT-PCR: DNA contamination, one vs two step - RNA work (Dec/22/2010 )
check this link for images of RNA run on an agarose gel:
http://www.ambion.com/techlib/append/supp/rna_gel.html
If you can see the ribosomal bands and your RNA isn't smeared to the bottom, you definitely have good enough RNA for RT-PCR. Partially degraded RNA can be fine if you are amplifying 100-200bp fragments.
Since you are new to RT-PCR, I recommend starting with two-step as it is more reliable. If you know your primers work with one-step, you can go ahead with that.
-David C H-
David C H on Wed Dec 22 16:17:09 2010 said:
check this link for images of RNA run on an agarose gel:
http://www.ambion.com/techlib/append/supp/rna_gel.html
If you can see the ribosomal bands and your RNA isn't smeared to the bottom, you definitely have good enough RNA for RT-PCR. Partially degraded RNA can be fine if you are amplifying 100-200bp fragments.
Since you are new to RT-PCR, I recommend starting with two-step as it is more reliable. If you know your primers work with one-step, you can go ahead with that.
I'll take into consideration these notes,
Many thanks
-noyara-