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Do you need to use the GOI for absolute quantification in qRT-PCR?
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Iā€™m currently working on an experiment in my lab. We are trying to measure the expression of a gene of interest (GOI) using qRT-PCR. Iā€™m not a big fan of using comparative quantification for this specific project and so I am trying to understand how to properly generate a standard curve.

Would I need to utilize the GOI for the standard curve or could I purchase some DNA or RNA at a known concentration that is not related to my GOI to generate the curve for absolute quantification?

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1 year ago