For dispense volumes 40 L - 150 L
Costar half area EIA/RIA plates (Costar, catalog #3690)
3. Certified pipettor (previously calibrated with dispense reagent) and tips for obtaining pathlength-vs.-volume calibration curve. (References 3,4 & 7-9 all contain procedures for gravimetric calibration of single pipettors taking into account the density of the liquid.)
OBTAINING A CALIBRATION CURVE
(Note: this procedure may be omitted if you have already generated a calibration curve)
Creating Plate sections for the calibration curve
A certified pipettor is used to dispense defined volumes of reagent (at least 8 replicates per volume) into the wells of a microplate. You can use a single microplate for the entire calibration curve. However, you should pipet, then immediately read, the set of wells for each given dispense volume, in order to minimize evaporation errors. You will therefore need to create a separate Plate section for each dispense volume, with the appropriate strip (i.e., the column in the microplate corresponding to the dispense volumes location) selected to be read.
Step 1 Select an empty Plate section, or create a new Plate section.
Step 2 Set up the Instrument Settings dialog box for the Plate section as shown in Figure 1. Select to perform an endpoint read at any wavelength. (SOFTmax PRO requires that you enter a UV/Vis wavelength at which to read, even though this measurement will not be used in calculating pathlength. Select PathCheck with Water Constant and deselect Pre-Read Plate. (Pre-reading is unnecessary because the UV/VIS measurement is not used in calculating pathlength.) Double click the Strips... button and select to read only the first column of the microplate.