We recently have purchased a copy number assay from Life Technologies. The assay performed well, as usually they have guaranteed the performance only when using their reagents, eg. their qPCR mastemix, so we did so. However I was interested how it performs with other mixes, since experiences suggested it will work well with another good mix as well. So let´s see the results:
This is a dilution series of a template that was determined as diploid with CYP2D6. We have determined 5 ng/ul as template concentration with a Nanodrop instrument and I made a 2-fold dilution series. The correlation coefficient is quite bad, the same is true for the variation of efficiencies. Since the template quantity is the same for the target and reference as well, it does not explain the difference for the undiluted sample.
This is exactly the same run with the same templates with PCRBIO´s probes mix. It is clearly visible that the correlation coefficients are absolutely acceptable and the efficiencies are practically the same.
The conclusion is that it is easy to find an alternate mastermix to replace Life Tech´s own mix that is cheaper and provides better results. Also I have tried in another run with its own protocol and PCRBIO´s run was 60 mins while the ABI runs for 90 mins.
Happy qPCR to everyone 🙂