How Do You Normalize Viral Gene Expression in Infection Studies?

Virology
1

Hi There

We are comparing viral gene expression across infected cell samples. Which reference genes or normalization strategies have given you the most reliable results?

Regards

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2

Hi Lee

We normally use a housekeeping gene for normalization and calculate relative expression using ΔΔCt.

I think the most important part is choosing a reference gene that doesn’t change after infection, because that can really affect the results.”

i hope it helps. Thanks

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3

Hi Both

In our infection experiments, we found that GAPDH was not always stable, particularly at higher infection levels. We therefore tested a few housekeeping genes first and used the most stable one for normalization. We also kept RNA input consistent and included mock-infected controls.

For relative expression, we used the ΔΔCt approach and confirmed that the PCR efficiencies were comparable before making the final comparisons.

Thnx.

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