Despite the advancement of new molecular biology techniques like ddPCR or RNAseq, quantitative PCR (qPCR) is still a golden standard for the quantification of nucleic acids, with applications in diagnostics and research. The method is based on real-time monitoring of amplification of the target DNA allowing its quantification (see basics). Yet, for the method to be quantitative, it has to be done properly.
Since we live in the digital era, why shouldn’t a popular method like PCR go digital as well? What is digital PCR (dPCR) and how it is different in comparison to PCR and qPCR, has been explained in the webinar “Why go Digital in qPCR”.
Price is often one of the most important factors when considering a purchase of a new qPCR thermal cycler. One reason for this is that it obviously has a direct and immediate impact on your budget.
Can we trust your published qPCR data?
The pressure to publish is causing shortcuts which result in misleading papers being published.
Will ddPCR replace qPCR technology?
Amplification efficiency higher than 100% would mean that in each qPCR cycle, more than two copies of the target sequence have been generated.
Is this possible?
How to present the ratio data?
Which are the most probable reasons for a false positive result and how to address the issue?
Researchers often wonder which statistical tests they should use with their qPCR data. Most frequently these questions arise when analysing gene expression data.