Efficiency of the m-PCR
The efficiency of the m-PCR was evaluated for the simultaneous detection of the target pathogenic microorganisms. The m-PCR was developed by using a mixture of DNA extracts from the five selected pathogens and compared
using the same test performed on the individual pathogens (Fig. 3). The results showed that the efficiency was not influenced when detecting mixtures of multiple target pathogens in comparison with the individual pathogens.
Optimization of the m-PCR
Optimization of the number of PCR cycles is shown in the image (not shown). Four of the five genomic DNA samples
were proved to provide better templates. The results also showed a gradual increase in the yield of each band with
increasing number of cycles. The greatest increase in the amount of products was found at about 32 cycles, while 35
cycles were optimal for the amplification of the target gene sequences. The findings for the optimization of annealing temperature are shown in the image (not shown). It can be seen that lowering the annealing temperature by 5 C was