ikekim at merle.acns.nwu.edu.
Fri Aug 25 09:29:12 EST 1995
Organization: Northwestern University
Reply-To: ikekim at merle.acns.nwu.edu
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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.
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