I’ve always struggled with this, but what is a good gene to use for normalizing Northerns and qRT-PCR results throughout development? I know some people use actin, but I’ve heard other people say this is not a good control to use because the expression of actin changes throughout development. Ideally I want to use a control gene that stays constant throughout development, so I can determine if the expression of my gene of interest changes throughout development.
I haven’t used these myself (our lab uses tubulin), but you may find something useful in this study:
Selection and validation of a set of reliable reference genes for quantitative sod gene expression analysis in C. elegans. Hoogewijs D, Houthoofd K, Matthijssens F, Vandesompele J, Vanfleteren JR. BMC Mol Biol. 2008 Jan 22;9:9.
There is also an older thread that might be helpful: http://www.wormbase.org/forums/index.php?topic=179.0
I have always used two different normalization controls (actin and ama-1) after reading the paper recommended by “Steve”.
Alternatively, you could mine the available microarray datasets for genes with stable expression throughout development, then test a few to determine which is the best behaved under your assay conditions.
Good luck,
Harold
ama-1, actin or eft-2 worm well. we have seen actin a little lower in embryos