Biotek EL800, Руководство оператора

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Chapter 4: Instrument Qualification
BioTek Instruments, Inc.
Plate Preparation:
5 Pipette 200 µl of the concentrated solution from tube 1 into each well of
the first column, A1 to H1, of the microplate.
6 Pipette 200 µl from each of the remaining tubes into the wells of the
corresponding column of the microplate (tube 2 into wells A2 to H2,
etc.).
Note: The choice of dilutions and the absorbance of the original
solution can be varied. Use Table 3 as a model for calculating the
expected absorbances of a series of dilutions, given a different
absorbance of the original solution.
After pipetting the diluted test solution into the microplate and before
reading the plate, we strongly recommend shaking the plate for four
minutes. This will allow any air bubbles in the solution to settle and the
meniscus to stabilize. If a plate shaker is not available, wait 20
minutes after pipetting the diluted test solution before reading the
plate.
Read Plate
1 Read the microplate prepared above five times using Normal Read
Mode dual wavelength at 450 nm with 630 nm as the blank.
Note: Do not discard the plate; you will use it for the Alignment Test.
2
Print the raw data or export it to an Excel spreadsheet using Gen5™,
KC4™, or KCjunior™.
If you are exporting the data to an Excel spreadsheet, perform the
calculations described below and on the following page, and keep the
spreadsheet for future tests.
Linearity Test:
3 Calculate the mean absorbance for each well, and average the means for
each concentration.
4 Perform a regression analysis on the data to evaluate linearity.
For example, using Microsoft
®
Excel:
In a spreadsheet, create two columns labeled X and Y. Enter the actual
absorbance values in column X. Enter the expected absorbance values in
column Y.