Strategy for finding RT-PCR housekeeping gene

Paul K. Farmer pfarmer at emory.edu
Mon Apr 26 12:45:42 EST 2004


We're performing RT-PCR on RNA extracted from cultured cells treated
with several potent reagents (alone and in combination with one
another). Our gene-specific primers work great (good specificity,
efficiency, reproducibility, etc). Unfortunately, our treatment effects
are modest two-fold changes. Furthermore, we have not been able to
identify any primers corresponding to a "housekeeping gene" that
provides constant mRNA levels in response to cell treatments. We've
tried several primer pairs used by others in different systems, but the
abundance of these transcripts have also varied with cell treatments. Is
using microarray analysis the best way to identify potential
housekeeping genes whose mRNA levels remain constant? This seems an
expensive and time consuming task, but so does the trial and error
approach involving testing primer sets used by others.

Paul K. Farmer, Ph.D.
Emory University Department of Medicine
Division of Endocrinology
WMB Building, Room 1327
1639 Pierce Drive
Atlanta, Georgia 30322
Laboratory: (404) 727-1393
FAX:  (404) 727-1300
Pager:  (800) 739-3013
Email:  pfarmer at emory.edu


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