What is the meaning of ΔΔCt in quantitative PCR and how is it calculated?

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The ΔΔCt value is a critical metric in quantitative PCR (qPCR) used to compare gene expression levels between different samples. It is calculated by taking the difference between the Ct (threshold cycle) values of the target gene and a reference gene in both the experimental and control groups. The formula 2^-ΔΔCt is commonly used to determine the relative expression levels, assuming that the PCR efficiency is 100%. This means that each cycle doubles the amount of DNA, which is a key assumption for accurate interpretation of the results. The discussion highlights confusion around the terminology and the underlying assumptions of the ΔΔCt method, emphasizing the need for clarity in understanding these concepts in molecular biology.
Bio-student
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I'm a little confused about what the ΔΔCt value means and how its calculated - could anyone verify?

Also, I'm reading lecture notes that say "2^ΔΔCt assumes efficiency is 100%". I'm really baffled by this and would appreciate any guidance.
 
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