# normalizing between qPCR runs - (Nov/09/2008 )

Hi,

I'm trying to normalize data for an experiment that I repeated, but I'm not sure how to normalize data for the two independent experiments. For each experiment, I normalized each datapoint to GAPDH, and each of those values was normalized to the first control time point (arbitrary unit = 1). Since each experiment's arbitrary unit 1 is not the same value when un-normalized, how would I do statistics to pool together the two independent experiments?

Thank you.

-BottegaVeneta-

I guess you should do statistics on replicates of each experiment alone. If you pool the results of two independent experiments you may get falsely positive or negative data.

-yobou-

QUOTE (BottegaVeneta @ Nov 9 2008, 09:05 PM)

Hi,

I'm trying to normalize data for an experiment that I repeated, but I'm not sure how to normalize data for the two independent experiments. For each experiment, I normalized each datapoint to GAPDH, and each of those values was normalized to the first control time point (arbitrary unit = 1). Since each experiment's arbitrary unit 1 is not the same value when un-normalized, how would I do statistics to pool together the two independent experiments?

Thank you.

I'm trying to normalize data for an experiment that I repeated, but I'm not sure how to normalize data for the two independent experiments. For each experiment, I normalized each datapoint to GAPDH, and each of those values was normalized to the first control time point (arbitrary unit = 1). Since each experiment's arbitrary unit 1 is not the same value when un-normalized, how would I do statistics to pool together the two independent experiments?

Thank you.

If I get it right, what you meant, don't pool. Normalise each sample to its control on the same plate and get the final relative normalised ratio (fold increase/decrease from the calibrator). You can compare the ratio between the two plates then (calibrator in both as a 1), because the differences will cancel out.

-Trof-