What is the acceptable coefficient of variance or SD in the CT values of reference gene in biological replicates (in treated and non treated samples)?

01 November 2022 1 4K Report

what is the threshold values for selecting or rejecting the gene as reference gene? how much intergroup variability can be tolerated?

Katie A S Burnette

That's not a valid concern. If you have different amounts of starting materials, you would expect to have different Ct values in your reference gene between BIOLOGICAL samples. That's the entire point of including this control.

What you should be worried about are TECHNICAL replicates. R squared values should be as close to 0.99 as possible to publish.

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