3.2 LAMP using genomic DNA
Figure 5 presents the sensitivity and specificity of the LAMP reactions using yaiO2 primers and genomic DNA of bacterial strains. We set 3 concentrations with 1% dilution with 5.68x1010, 50% dilution with 2.87x1011, 100% with 5.34x1011 copy numbers for E. coli and 1% dilution with 6.82x1010, 50% dilution with 2.72x1011, 100% with 5.74x1011 copy numbers for P. aeruginosa (100% dilution ≈ bacterial culture of a mean size colony). We conducted the LAMP assay with the same parameters used for the Colony-LAMP assay. Figure 5ademonstrates the LAMP reactions using genomic DNA in the PDMS wells. The first row of the PDMS slab were pink, which presents the negative controls both for E. coli and P. aeruginosa strains. All the wells belong to the dilutions of P. aeruginosa genomic DNA were also negative. The rest of the wells were in yellow color and containing the serial dilutions of the E. coli genomic DNA in triplicate. We evaluated the results of the LAMP assay using electrophoresis, Figure 5b . We obtained the ladder-like patterns for all the dilutions of E. coli genomic DNA in the gel electrophoresis. We did not observe similar bands for the genomic DNA dilutions of P. aeruginosa , Figure 5b . LAMP assay using genomic DNA also confirmed the specificity, sensitivity of the LAMP method and robustness of the LAMP platform.