Calibrator and standard

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islam
islam's picture
Calibrator and standard

Dear all,
I would be very happy to learn about the standard and calibrator of  Real time PCR. During PCR we usually use calibrator and standard. For example , say I would like to amplify alpha receptor mRNA, for which I need standard alpha mRNA. We use calibratore to normalize between plate to plate. My question is there any certain roles for the strenght diffrences between Calibrator and standard sample. If so how many times strenght  differences  between this two preparation  is better and why?

Thanking you

Regards
Islam 

Ivan Delgado
Ivan Delgado's picture
 

 
Hi Islam,

Unfortunately the nomenclature in real time PCR is not very consistent, so while some people use "calibrator" (introduced by Applied Biosystems) others use "reference". Here is what I can tell you: 

- A standard is a sample of known concentration. A standard is typically only needed if you are running an absolute quantification experiment, which basically means you run a standard curve using your standard sample. You can then use this standard curve to quantify the expression of your gene(s) of interest in unknown sample(s).

- A calibrator, or reference, is typically a sample for which you do not really need to know the concentration. This is because this reference sample is most commonly used in relative quantification experiments, where the expression of your gene(s) of interest in unknown samples are measured relative to their expression in this reference sample. 

Hope this helps. 

islam
islam's picture
Dear Ivan,

Dear Ivan,
Thank you very much for your suggestion. May I have your mail Id, please?

Regard
Islam

Ivan Delgado
Ivan Delgado's picture
 

 
Sure. I sent it to you by PM. Let me know if you do not get it.