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Quality control of RNA (quantity, quality and purity)
Reverse transcription
Aspects to consider early on
qPCR primer design
The qPCR run
Optimizing qPCR assays
Primer efficiencies
Reference/housekeeping genes
Calculating relative gene expression
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Data analysis options (4:26)
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The Delta-Delta Ct method (3:45)
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Performing the Delta-Delta Ct method using Microsoft Excel (6:12)
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The Pfaffl method (2:00)
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Performing the Pfaffl method using Microsoft Excel (6:53)
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Relative gene expressions with multiple reference genes (Vandesompele method) (2:28)
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Performing the Vandesompele method using Microsoft Excel (6:57)
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The relative quantities method (2:01)
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Performing relative quantities method in Microsoft Excel (3:01)
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Always log-transform your data before statistical analysis (6:05)
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End of unit quiz
Additional resources
