In the multiplex PCR system, preferential amplification
of one target sequence over another frequently occurs,
which could lead to uneven amplification products. InThe results showed that species specific for 16SrRNA
gene of A. hydrophila could not be detected when Mg2+
concentration was 1.5 mM. The amplification efficiencies
improved by increasing Mg2+ concentration to 3.5 mM.
Nevertheless, excessive Mg2+ concentration resulted in
the nonspecific amplification and decreased specificity.
Consequently, Mg2+ concentration of 3.0 mM was optimum
for multiplex PCR reaction (Figure 4)