To illustrate its predictive power this method was used in multiplex PCR design to successfully generate four resequencing assays containing up to 126 primers with no observable primer-primer amplification artefacts.
To detect multiple target bacterial species simultaneously, gene specific primers designed to target the proteases genes of Acinetobacter ssp., P. fluorescens, Aeromonas hydrophila and Serratia marcescens were used in multiplex PCR.
For Bacteria, the primers (Table S1) were used in multiplex [35].
Also, DNA primers encoded by MCTs have notably been used in multiplex PCR microbe detection assays; positive test results were verified by real time PCR [17], [18].
The DBSS have also been used in multiplex analysis of inflammatory markers [17] and in 2008, the Jeffrey Modell Foundation and the State of Wisconsin launched a DBSS based newborn screening program for severe combined immunodeficiency [18].
The breakpoint junction and control primers and probes were used in multiplex within one reaction.
Utility of loci to be used in multiplex PCR reactions relates to amplicon size, dimer formation, stutter noise, annealing temperature, and primer complexity.
Template DNA from stool isolates was used in multiplex PCR to investigate the presence of toxin genes, as described elsewhere, with minor modifications [ 12].
The consensus primers (VP7-F/VP7-R and con2/con3) were used in multiplex first round reactions and type-specific primers (G1 G4, G8 G10 and P[4], P[6], P[8] P[11]) were used in the second round reactions.
