quantitative RT-PCR

[Isaac Kim]

Organization: Northwestern University
Reply-To: ikekim at merle.acns.nwu.edu
X-Newsreader: WinVN 0.91.6


I have used quantitative RT-PCR for past publications. I have had no 
problems with the reviewers. The main problem with RT-PCR is that 
different primers have different efficiency of amplification. If you 
choose the conditions of the standard correctly, RT-PCR can be more 
accurate than Northern blot analysis, in my experience. Even more 
interestingly, a recent manuscript that I submitted to a reputable 
journal, the reviewers' response was that I should use quantitative 
RT-PCR over that of Northern blot analysis for all my studies as the 
control for Northern, GAPDH, showed variations in the level. I would like 
to direct you to two articles regarding this matter. 
Siebert PD et al. 1992. Competitive PCR. Nature 359:557-558
Forster E. 1994. Rapid generation of internal standards for competitive 
PCR by low-stringency primer annealing.

Isaac.