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Methylene blue can adsorb on ss-DNA and increase its concentration whereas the part of the DNA probe involved in hybridization near the detection limit is very low so that higher number of binding sites would result in lower part of the DNA probe converted to ds-DNA and hence in lower relative decay of the Methylene blue signal. The calculations of LOD could confirm the authors suggestion but they did not take into account the standard deviation of the signal. It was determined as 3% for upper quantification limit of target DNA. This means that at least 10% decay of the Methylene blue signal (S/N=3) would correspond to LOD. As could be seen from the Fig. 8 and Fig. 9, 2 microA shift of the signals corresponds to the target DNA concentration of approx. 10-16 mol/L, 100 times higher than LOD declared. The error bars shown in Fig.9A are much lower than those expected from the S.D. announced. The authors did not provide the description of the Methylene blue signal in blank experiments and this complicates the assessment of the metrological characteristics of the DNA sensor.
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