Primers were tested for optimal concentration in triplicate or quadruplicate in combinations of final concentrations of 100, 300, 600, and 900 nM; the probe was tested in triplicate at final concentrations of 50, 100, 200, and 300 nM.
Water samples from experiment 1 were tested for a third time in triplicate.
All samples were tested for degradation and quantified in triplicate using fluorimetry, employing PicoGreen® dsDNA quantitation reagent (Cambridge Bioscience, UK).
Analyte stability evaluation was also included in the experimental layout: Odd-chain standards were spiked in naïve human plasma to a final 500 nM concentration (triplicate samples) and tested for stability in both short-term (4 and 25 °C for 6 h) and long-term conditions (9 days at −20 °C).
Data were obtained in triplicate and expressed as mean and tested for significance by P < 0.05.
The synthesized silver nanoparticles were tested (triplicates) for minimum inhibitory concentration by microtiter broth dilution method [ 32].
Randomly selected transformed cell colonies were tested in triplicates for anchorage-independent growth in soft agar by previously described procedure [16].
All samples were tested in triplicates for each genotype and the results represent averages of individual experiments.
Each strain/concentration combination was tested in triplicate for each assay, and the entire assay was repeated three times for each of the strains of H. ducreyi tested, as well as for all control organisms.
All samples ("aged" for 14 days) were tested in triplicates.
