( the tip and the number of mixes was v...)Serial Dilution with the Bravo Liquid Handling Platform

the tip, and the number of mixes was varied from 3-20 cycles. Table 1 shows the resulting CV of each column for each mix cycle protocol. The average precision (averaging CVs for columns 1-10) shows that the precision improves asymptotically as the number of mix cycles is increased (Figure 2). Three mixes yielded an average CV of 11.8%, while 20 mixes had a considerably better CV of 1.7%. The precision in all cases worsens as the serial dilution proceeds across the plate; this is expected as the error in the earlier columns is propagated with each transfer.

In addition, the accuracy ratio improves as the number of mix cycles increases (Figure 3). The accuracy ratio is an average of the concentration of the diluted column compared to the previous columna perfect serial dilution would have an accuracy ratio of 1:2.00 across the entire plate. The accuracy ratio of the plate improves with more mix cycles, varying from 1:1.85 to 1:2.01. This is expected as the fluorescein would be more equally distributed in the well the more the well contents are mixed, ensuring that transfers are made from a uniformly mixed solution.

While the precision and accuracy with 20 mix cycles is very close to a perfect serial dilution, it also undoubtedly takes too much time for an automated process. The 20 mix cycle protocol takes 20 minutes to perform, while a 3 mix cycle protocol takes less than six minutes. Efforts were then focused on the factors that can improve the 3 mix cycle protocol to produce accuracy and precision results that were closer to the achieved 20 mix cycle protocol.

Mix Height
The mix height was adjusted in order to determine the effect of distributing the liquid at a different location in the well. As the mix height was increased, the average precision improved. Figure 4 shows that at a height of 3mm, the average pre
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