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.