Nucleic acid analysis has become increasingly important in a
variety of applications, such as the genotyping of individuals, the
detection of infectious diseases, tissue typing for histocompatability,
identifying individuals in forensic diagnosis, paternity
testing, and monitoring the genetic make-up of plants and animals
in agricultural breeding programs [1]. Techniques based on
polymerase chain reaction (PCR) provide a powerful tool for the
amplification of minute amounts of initial target sequences. Most
PCR protocols involve reactions that amplify a single target.
Multiplex PCR is a variation of the conventional technique in
which two or more targets are simultaneously amplified in the
same reaction. This approach has the potential for greater
reliability, flexibility, and cost reduction. As far as we know,
nine-target multiplex PCR method has been reported to
simultaneously detect eight maize lines as well as the endogenous
Zein gene in a single reaction tube [2], which contains the most
targets in reported multiplex-PCR methods.