Overview: Multiplex PCR is a variant of PCR which enabling simultaneous amplification of many targets of interest in one reaction by using more than one pair of primers. Since its first description in 1988 by Chamberlain et al, this method has been applied in many areas of DNA testing, including analyses of deletions, mutations, and polymorphisms, or quantitative assays and reverse transcription PCR. Typically, it is used for genotyping applications where simultaneous analysis of multiple markers is required, detection of pathogens or genetically modified organisms (GMOs), or for microsatellite analyses. Multiplex assays can be tedious and time-consuming to establish, requiring lengthy optimization procedures.