How to design your qPCR experiment so that it conforms to the MIQE guidelines
There have recently been some high profile retractions of scientific papers that have reported data which turns out to be artifacts of poorly designed experiments. The MIQE guidelines is a daunting list of 85 items that need to be addressed before a reviewer will accept your qPCR results as valid.
In the PDF below, the essential elements of sample preparation and experimental design are outlined as a practical guide to meeting the MIQE guidelines. If you use the approach followed in this guide, you will save countless hours of effort trying to figure everything out on your own. Sean Taylor, Michael Wakem, and Greg Dijkman have contributed their many years of qPCR experience, and when you follow this guide, your gene expression experiment will meet the MIQE guidelines.
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This is a reprint of the article published in Methods: April 2010
If you are not already testing your RNA quality then you should consider acquiring an instrument like the Experion to validate the quality of your RNA. If you do not already have a Bioanalyzer or an Experion, you will need to run your RNA samples out on a gel.
Here is a real case study of data that gave inconclusive results due to many of the MIQE recommendations being ignored. Have a look at the data after the experiment was repeated using the advice from practical guide to MIQE guidelines.
Open the MIQE case study pdf here.