Dear researchers;
A question arose for me about Real-Time analysis. I would appreciate you to comment about the question.
For expression analysis we need a housekeeping gene to normalize Ct values. When calculating expression ratio of two genes like Bax/bcl-2 in different samples, do we still need to normalize Ct values against housekeeping gene?
Hi. there are many method to calculate the gene expression, the common one is Livak equation.
in this method you have the values of CT of the housekeeping gene and the two genes you used then you calculate it as the following :-
1- ΔCt= Ct gene of interest – Normalization Factor
2- ΔΔCt= ΔCt sample A – ΔCt calibrator .
3- Fold Change = 2-ΔΔCt and this is the gene expression
i suggest to you this ppt for me
Presentation Basic of quantitative real time PCR
and waiting any other question from you
I recommend the use of at least one well-chosen reference gene for normalization as it takes care of potential differences of overall DNA or RNA concentration or integrity among your samples. Thanks
If you want to publish your data, you will need to show a house-keeping gene. Article The MIQE Guidelines: minimum Information for Publication of ...
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