hi im a new researcher. i did qPCR recently, looking at gene expression change in certain genes. basically in a plate, i ran 3 biological repeat controls, and 3 biological repeat mechanically compressed cells.
i expose compression onto cells for 6 hours, and for two genes i looked so far, i see a fold change of about 0.89 and 1.2. And this is consistent within each controls and compression, that is its quite precise but upon running t test, it is not significant.
i was wondering if i increase compression from 6 hrs to 16 hrs, will i 'enhance' the effects a bit more such that any difference will be significant?
Well, i think you can get a try, set a series gradient of treatment. However, before that, it's better to check your reaction details such as primers' specificity and efficiency etc. Make sure they are suitable for your qPCR reaction. Good luck!
The best way to test this would be to perform a time course. You could also add in a positive control, depending on the genes you are looking at. This would confirm if your conditions are optimal for detecting changes in gene levels.
Dear Iwan: Do not forget that you have to use several standard genes as references, and test that they do not change (low variance) with you treatment.
See this work by Vandesompele et al. http://genomebiology.com/2002/3/7/research/0034.1
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