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Troubleshooting qPCR


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#1 EMSA1983



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Posted 21 April 2012 - 08:45 AM

I have been doing qPCR to examine gene expression changes in chondrocytes in a 0-20days timecourse. I see that some genes are consistently upregulated during the timecourse but the problem is I am not getting reproducibility in the various runs. I have done several batches so far and in every batch I see that there is an upregulation but the numbers are not consistent. So when I plot the error bars, they are really high. I always normalize the expression against housekeeping gene GAPDH. Not sure what's going wrong. Any help will be appreciated! Thanks..

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