Smart PCR leading to improved DNA profiles

Our ultimate aim is to develop a smart PCR system that can monitor amplification in real-time and amend PCR conditions on-the-fly to obtain better quality DNA profiles from challenging samples. For such a system to exist, a method is needed to monitor the progress of an amplification reaction in real-time without compromising the quality of the DNA profile. This study used a combination of GlobalFiler^TM and the Investigator Quantiplex Pro RGQ Kit for PCR and found the combination-multiplex amplified the highly polymorphic regions (STRs) used for DNA profiling and quantification regions used for DNA quantification in a single reaction. By using this combination, it is now possible to amplify STRs for DNA profiling whilst also monitoring the kinetics of the amplification reaction in real-time. The combination of these reactions was performed on an open-source qPCR machine to allow the real-time fluorescence data to be collected before DNA profiles were generated using capillary electrophoresis.