Biological sets

[sylvia.c.wong]

Hello everyone!

I am fairly new to the GenEx program. If I have cDNA samples from different number of animals (for an ex. One cDNA tube is from two animals, and another tube of cDNA is from three animals) and then used the templates in qPCR in triplicates, how can I normalize my data? Thanks!

-S

[Mikael Kubista]

This question is rather biological than technical. To approach it, what kind of variation are you interested to measure and what kind of variation do you like to remove with the normalization?

Nevertheless, the standard approach of comparing expression of candidate reference genes in representative samples with the Normfinder is generally valid. You may also consider normalizating to total RNA.

Good luck!

[sylvia.c.wong]

Thank you for your reply. I want to correct the different number of treated animals I am using vs. untreated (control). I have one tube of cDNA synthesized from RNA from 3 treated animals and one tube of cDNA synthesized from RNA from 2 untreated animals. I want to correct this variation. Do I need to correct this? How can I do that with GenEx? Thank you!

-S

[Mikael Kubista]

Unfortunately, if the samples of each kind have been merged prior to analysis, variation can no longer be estimated. It is better to analyze all samples separately. In GenEx groups of each kind of samples are created in Data Manager and then compared using appropriate statistics (t-test usually). GenEx then accounts for the different number of samples in the two groups.

Good luck