Relative transcript quantification by Quantitative PCR: Roughly right or precisely wrong?
Marine Genome Biology, Institute of Marine Research, N-5817 Bergen, Norway
BMC Molecular Biology 2005, 6:10 doi:10.1186/1471-2199-6-10Published: 26 April 2005
When estimating relative transcript abundances by quantitative real-time PCR (Q-PCR) we found that the results can vary dramatically depending on the method chosen for data analysis.
The results emphasise the importance of being cautious when analysing Q-PCR data and indicate that uncritical routine application of an analytical method will eventually result in incorrect conclusions. We do not know the extent of, or have a universal solution to this problem. However, we strongly recommend caution when analysing Q-PCR results e.g. by using two or more analytical approaches to validate conclusions. In our view a common effort should be made to standardise methods for analysis and validation of Q-PCR results.