Just a comment on David L's comments on housekeeping genes: David Leavesley wrote: <lots of good stuff which I have snipped> > Beta 2 microglobulin was recently suggested to me on the basis that ALL > cells express it (even my meso/epithelial cells where it too is a rare > message). I had intended to use b2m as a housekeeping gene for my early porcine embryos. I am collecting them just around the time they start producing our genes of interest - IFN-gamma and IFN-delta (a type I unique to the porcine trophoblast). In our (admittedly fumbling) hands, we got more reliable amplification of our IFNs than our so-called housekeeping gene. After further reading, we learned that b2m expression was INDUCED by IFN-gamma. (not a new finding but new to us!) Somewhere around this time, I reached the conclusion that there probably was no such thing as a housekeeping gene in a rapidly developing embryo! We eventually decided to try Ambion's QuantumRNA kit (based on 18s ribosomal RNA as an internal standard) for our semi- quantitative' RT-PCR. Can't comment on how well it works - this project is currently 'on hold' while we go back to more familiar frustrations! Heidi -- ** note new email address ** Heidi Engelhardt, Adjunct Professor Department of Animal & Poultry Science University of Guelph Guelph, Ontario CANADA N1G 2W1 email: hengelha@uoguelph.ca phone: (519) 824-4120, ext. 3658 fax: (519) 767-0573
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