gene of interest or housekeeping gene shift...? - (Sep/29/2006 )
i have just run a real-time pcr and the data i believe id untrustworthy... i ran 4 control samples and 4 treatment samples (in quadruplicate) and found that between the 2 groups, the gene of interest had a similar Ct value but the housekeeping gene average Ct was reduced in the treatmenmt group. when comparing ddCt this make it look as if the treatment has increased the gene of interest... how unreliable is this data?
You need to be able to show good consistency in your housekeeping gene. There should be minimal fluctuation otherwise it will be difficult to get it past referees for publication.
what's your HK? perhaps you need to switch? some are sensitive to certain treatments
its ok, it was a shitty run... repeated the HK gene and was same as control.... thanks anyway guys, keep up the posting!