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

of 50μL of degassed deionized water. The deionized water is degassed by bubbling helium through the water for a period of 15 minutes.

Qualification of a Reagent Dispenser

In our testing for a "liquid to liquid dispense", a Bulk Reagent Dispenser on a Sciclone Liquid Handler is used to dispense 50μL of degassed water into the plates. Qualification of the accuracy of this dispense is performed gravimetrically.

Calibration of the Plate Reader

To calibrate the plate reader, 75μL of degassed deionized water is pipetted into a 384 well polystyrene plate (Nunc #265196). To this plate, 1μL of the Tartrazine solution is manually added to each of 10 wells by successive 1μL dispensing from a filled 10μL Hamilton micro syringe. After manually mixing with multiple aspirate-dispense cycles using a hand pipetter, the wells were read for absorbance reader. Similar procedures were done for the 0.5μL and 2μL plates. The average absorbance reading for these manual standards was used in a single point calibration routine. This assumes a proportional relationship between the volume of dye in the well and the well reading.

Setting up the Sciclone

In the Sciclone software, a layout is created with two reservoirs, (Nunc #242811), and five 384 Well, clear, non-treated plates (Nunc #265196). Prior to running the Sciclone protocols, the accuracy of the Sciclone is verified. Using the values measured above in Calibration of the Plate Reader, the absorbance values obtained are compared to those obtained using the Sciclone dispense. To achieve the desired accuracy, under the "Liquids" tab, a Liquid called "LDMSO" is created and given a first order calibration value of 0.94. This same value (0.94) is used with all three volumes described in this note.

Running the Sciclone Protocols


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