I have a problem in the representation of Real-Time PCR data. I analyzed the target (trypsin, chymotrypsin and elastase) genes expression in various independent treatments, i.e. Insects fed on three different diets. The target genes expression was quantified by relative expression analysis (normalized with stably expressed housekeeping gene). I want to present the data to show the relative expression levels of target genes but not normalised to first diet as one, i.e. If I normalize with first diet, the expression of all target genes become =1. I don’t want to represent in this way since this does not provide absolute expression levels of target genes. Therefore, please kindly help me how to represent the data to show absolute expression levels in the insects fed on different diets.
Please be more specific. I agree with last commentator. Question a bit ambiguous
I am very much clear about question as I am doing a lot of relative gene expression on real time PCR and faced same problem. it is very sample when we use term of Relative expression then we have to select one as control and we observe the effect of other treatment on gene expression of target genes ig increase than it shows fold increase as compare to value 1 mean 2 or 3 folds increase after normalizing with housekeeping like GAPDH or Actin. and in case of increase we take it as control 100 and decrease in value as percentage decrease.....
If I understood correctly there are several options:
1. You can use the formula 2 power -delta Ct, in this case you just normalize to housekeeping gene, and not to "control" sample. So can get rid of the 1 value, but the data will be still normalized and relative.
2. If you want to do quantitative, or absolute you have to order all your genes of interest and run serial dilution a do standard curve.
3. If you have possibility to run digital droplet PCR ( ddPCR) this will solve all your problems.
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