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384 Well Pipetting Precision and Accuracy Testing Using the Sciclone Advanced Liquid Handler

The Sciclone is programmed to aspirate a volume of tartrazine solution. After dipping the cannulas into a rinse solution to remove any residual external tartrazine solution, they are dispensed into a clear bottom plate containing water for the "liquid to liquid" dispense. The procedure dispensed into 5 plates sequentially. Apart from appropriate changes in volumes, the same protocol is used for the 0.5μL and 2μL tests.

Below is an example of a Sciclone protocol for a 1.0μL dispense:

Mixing the Plates

After pipetting the 5 plates, an extra 25μL of DI water was added, again using a separate qualified reagent dispenser as described in MATERIALS AND METHODS to aid in the mixing of dissimilar liquids in the well. Next, the plates were given 15 shake cycles on a Molecular Devices V-max reader and allowed to set for 15 to 30 minutes before they were read.

Reading the Microplates

Plates were read for absorbance at 450nm. The raw results were exported to ExcelTM and CVs were calculated for each volume.

Results

The three tables to the right show the precision and accuracy observed in this testing. The accuracy is shown as the average volume dispensed. The precision is given in CV for the individual plates and the 5 plate average at each volume. The values in the tables were imported from Excel sheets.

Discussion

The experimental data demonstrates precise pipetting performance that can be attained using the Sciclone ALH. While Factory Release Dispensing Specifications for the Low Volume Head are 8% CV for volumes of 2μL and below, and 3% CV for volumes of 2 to 25μL, in this work we achieved CV's that are significantly better than the specifications. Interestingly, the first plate dispensing 0.
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