quantitative RT-PCR

Richard R. Hardy hardy at mighty.fccc.edu
Fri Aug 25 14:33:01 EST 1995

In article <41kmno$84 at news.acns.nwu.edu>, ikekim at merle.acns.nwu.edu.
(Isaac Kim) wrote:

>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.
And the second reference is...
Richard R. Hardy                       Member, Institute for Cancer Research           
Fox Chase Cancer Center                Tel:    (215) 728-2463
7701 Burholme Ave.                     FAX:    (215) 728-2412
Philadelphia, PA 19111                 E-MAIL: R_HARDY at fccc.edu

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