I'm performing RT-PCR analyses using actin, 36b4 and GAPDH as housekeeping. The samples are all concentrate around 10 ng/µL and the ratios 260/280 and 230/260 are in the range. Both controls and treated-samples show Ct for the housekeeping that differ in 10 Ct. The standard curve is correct, so I suppose that is not a mix problem nor a loading problem. Can anyone help me solving this problem?

Thanks in advance

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