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Reference Gene/HKG- every plate

qpcr reference gene

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#1 ryc



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Posted 02 March 2015 - 03:50 PM



Is it necessary to run reference genes/housekeeping genes on each plate? For example is would it be okay to run the reference genes on plate 1 and then compare target genes run on plates 2 & 3.


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#2 bob1


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Posted 04 March 2015 - 01:25 PM

You would need an inter-plate comparator to do this, otherwise you should run your HKG on every plate.

#3 Trof


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Posted 07 March 2015 - 03:30 PM

Actually that is not correct.

In relative quantification you only need all the samples (including calibrator) to run at the same plate. Different genes can be run separately, including the HKG, because the Cts among the genes are normalized anyway in the calculation.

Only exception is if you have so many samples that they don't fit one plate. In that case you need to include the calibrator on every plate and calculate the values from one plate compared to that specific calibrator.

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