While calculation of the fold change expression of a transgene/gene, the expression level of target gene is normalize by subtracting delta Ct of treated sample with delta Ct of control sample. Suppose, one wants to evaluate fold change expression of a transgene in transgenic plants exposed to 50, 100 or 200 mM NaCl stress. A native homolog of the transgene exists in the transgenic plant (eg. AtCBF3 & LeCBF3). In this case, Ct Value of which plant sample we should choose to normalize the expression of transgene. Ct of Non-transgenic plant exposed to respective salt stress (Control) or Ct of transgenic plants exposed to 0mM (control)?
My guess is that you can reference to nontransgenic plant in 0 mM condition. In that case, transgenic plant in 0 mM condition should not give you (i dont know if your promoter is leaky) variations compared to the reference. By the way, you will be referencing your data, so all your values will be relative. That will not change the overall results. Best.
I understand and suggest that you can compare your transgene line expression @ 0 mM with same line @ other conc of salt as mentioned by Margarit. At the same time non transgenic line should be compared with the transgenic lines for all the salt levels so you can compare variable level of your transgene expression in diifferent lines compare to non transgenic as well. This is more work but allows you to compare three way. As your non transgenic line used for transformation may have potential member of this gene family....
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA