Here we intend to use Digital PCR to qualify GMO reference material obtained from the Breeding companies for those GMO events which we cannot find Certified reference material on the market.
We get good results in cases where we have high % of GMO an a lot of amplifyable DNA. In the low range we have comparable high measurement uncertainty as with real time PCR. The Performance seems also to depend on the PCR Systems applied. This Counts for relative quantification. for absolute quantification we get high MU. But in the field of GMO we always use relative quantification.
Thank you very much for your contribution. Our problem is just because of those cases when we dont have certified reference materials to perform a relative quantification. We assume that our calibrants are 100%. So, We want to check if the seeds we´ve received from the Breeding Companies are really 100% (% GMO). In these cases, we need to perform an absolute quantification using digital PCR. Based on these calibrants we prepare a standard curve to perform a relative quantification of our samples. We already observe in our lab very high values of measurement uncertainty with dPCR quantifying in our limit of quantification compared to those obtained by Real-time PCR. I am reading the last paper published by your group. Here we use the Fluidigm system (Biomark).
You may contact the National Institure of Slovenia (Tanea Dreo; Jana Zel) and also the European Reference Laboratory for GM Food and Feed (Philippe Corbisier).