Part:BBa_K1565002:Experience

ColiAlert

This color variation is shown in the graphs of absorbance shown in figure 3 and 4. Figure 3 is an absorbance spectrum generated by readings from 300 to 680nm at pH 4 and 7. It is possible to see that the samples at neutral pH, having two absorption peaks, while acidic pH has only one, showing the transition of colors. In Figure 4 is possible to observe the change in absorbance obtained from samples of ColiAlert diluted in citrate buffer (0.1M) and phosphate buffer (0.1 M) at different pHs in readings of 500nM.

To try to show that ColiAlert signals the process of extracting nucleic acids correctly, we tested the extraction of nucleic acids only with blood, and blood together with ColiAlert. At the end of procedure, the resulting samples were quantified with NanoDrop (Results in Figure 5). It is possible to note the indirect evidence that in the presence of ColiAlert, the amount of DNA generated was more than twice as high. Figure 6 shows the variation in color and pH obtained after the lysis procedure and figure 7, 8 and 9 shows plates with ColiAlert. And figure 10 shows the process of reversible and irreversible change in coloration.

Figure 3. Curve of the absorption spectrum from 300 to 680nm.

Figure 4. The absorption spectrum of ColiAlert at different pHs (Readings at 500nM).

Figure 5. Quantification of extracted DNA (Nanodrop).

Figure 6. From left to right, ColiAlert after lysis solution, ColiAlert in acidic solution, and ColiAlert in basic solution.

Figure 10. Shows the process of reversible and irreversible change in coloration.

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