Relay-PCR™ is a new, simple, and robust method of multiplexing PCR. A single PCR run accomplishes two functionally separate reaction phases without any primer removal step, resulting in a significantly simplified, one step workflow. A single pair of common primers and a mixture of multiple pairs of specific primers are added to a genomic DNA sample in a single tube. Both a selection phase, where the genomic targets of interest are captured, and a library amplification phase, will occur during this single PCR reaction.
More importantly, the addition of the common primers induces a profound change to multiplexing PCR by limiting the role of specific primers to only the first two thermo cycles for target replication and enables an automatic switch to common primers in the remaining thermo cycles for library amplification. This arrangement eliminates a significant cause for amplicon to amplicon variations in end product due to priming efficiency variations among specific primers which would be exponentially amplified if used as amplification primers.
