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Problems with relative quantification in real time PCR

Problems with relative quantification in real time PCR - Molecular Biology Techniques

Problems with relative quantification in real time PCR - Molecular Biology Forum. Includes forums for common molecular biology techniques.


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Old 05-19-2012, 02:14 AM
Pipette Filler
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Default Problems with relative quantification in real time PCR



Hello everyone!
I am a new user of real time PCR an I really need some help to choose the best method to analyze my data.
In my experiments Im injecting an exogenous gene (GFP) in some animals (using a particular vector), and now I want to quantify the relative expression of this transgene in some tissues.
Initially, Im thinking to aplly the 2^-ddCt method but I have a problem... What should I use as a calibrator? The nontreated animals do not have transgene expression, so its not correct to use it as calibrator... Am I right??
What method should I use to analyse my data?
If anyone can help me, please let me know.
Thanks in advance,
LPS_C
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pcr , problems , quantification , real , real time pcr , relative , relative quantification , time


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