Exploring primer-template interactions using a deconstructed PCR methodology

As part of this study, 16 different experiments were conducted comparing the effects of PCR amplification method (TAS or DePCR) and annealing temperature (45°C or 55°C). Each experiment was a PCR amplification of synthetic DNA templates, ranging from a single template to a combination of up to 10 different templates. In some experiments, synthetic DNA templates were added to the PCR reaction mixture at equimolar concentration, while in others, each template was added at a different concentration. In addition to varying input templates, a set of 64 primers were used in different combinations to amplify the synthetic templates. In some reactions, only a single was used, while in others, various combinations of primers were used to amplifying synthetic DNA templates.