I am trying to quantify a miRNA of interest using the Ncode system. I would like to use GAP3DH/B-actin/18s RNA/ 4.5s RNA, etc. as an endogenous control because we dont yet have a good candidate miRNA that could act as a good control.
So I called their tech support and asked if after the poly-A tailing of miRNAs, the Universal RT primer would reverse transcribe mRNAs as well- so I could use the same RT product to run my real-time analysis for the miRNA and a control gene.
The person told me, the Univ RT primer is a oligo-dT with some minor modification. Hence she said it might not work the same as a normal oligo-dT (like in the superscript kits). BUT it does work for 5s RNA since it is small enough to get poly-A tailed. Although I believe her, I would REALLY like to know if someone else has used the Ncode system and quantified 4.5s RNA or other mRNA endogenous controls from the RT reaction products.
Any suggestions are greatly appreciated.
Edited by it_never_ends, 16 February 2009 - 12:12 PM.