Hi,
I have got Fold change for delta delta Ct values, but I cannot find the best statistics way to comparing among these genes from one side, and another side how can compare between stresses.
In attached, you find sample from my results.
Many thanks in advance
Saddam BABA
Hi Saddam,
If you inted to compare two groups of expression data, in your case 15 and 50% levels, I think that you could use a DDCt method implementing the qBASE analyzer software (https://www.biogazelle.com/qbaseplus) to Relative Expression calculation. Excel spreedSheet also works, however the primers efficiencies is not consided as occurs in qBASE Software.
For analyses, I suggest you to use one level as a calibrator (RQ= ~ 1.0) to calculate the FoldChange. Then to statistical analysis I have been used the REST Software (http://rest.gene-quantification.info/), this software uses a randomization test, with a customized boostrap (1000 as a default) parameter to estimate the significance of RQ diferences observed between two groups (named control and treatment).
Using two or three suitable references genes is also advisable approach. Searching the literature or validating your own reference genes (gene with no or low variation in response to the experimental conditions).
I hope that these tips could help you,
Best Regards,
Helder Anderson
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