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Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516
TM397 · Revised 12/16
www.promega.com
3.E. Analysis of Results (continued)
4.
Performing an Analysis (continued)
c. From the lower left side of the ‘Results’ screen, touch the
Analysis
button.
d. The ‘Templates’ screen will open, allowing you to select an existing analysis template file to apply to the
raw data. Touch the template that will be applied to the raw data. If there is not a preexisting analysis
template for this type of data, touch the
New
button in the bottom left corner of the screen to create a
new template as part of the analysis process.
If an existing analysis template is selected, the analysis screen will open with the raw data and template
values prepopulated.
If the
New
button is pressed, the analysis screen will open with raw data values prepopulated in the
plate map. Choose the curve fit type, well type designations, naming and concentration/dilution factor
following the steps of creating a template indicated in Creating a New Template, Steps f–n. This
template can be saved by touching the
Save Template
button in the bottom left corner of the screen.
When starting an analysis, the raw data from the plate read will be populated in the center of the wells
on the plate map. For the results of a single plate read, you can toggle whether the raw data is displayed
in the wells of the plate using the Raw On/Off button at the lower left side of the screen (Figure 56).
When analyzing ratiometric data from dual plate reads, the buttons allow you to toggle between Ratio
On/Off, Read 1 On/Off and Read 2 On/Off to display the ratiometric data or the individual raw data
from each plate read. Analysis of NanoBRET data modifies the buttons, allowing you to toggle between
Ratio On/Off, Donor On/Off and Acceptor On/Off in order to display the ratiometric BRET data or the
individual raw data from the donor or acceptor plate reads.
e. Only a single experimental data set can be analyzed by the software at a time. The nature of an
experimental data set is described below and is specific to the curve fit being applied to the data. Once
an analysis template has been applied to the raw data, make sure that only one experimental data set is
active. Select wells that will not be analyzed, and touch the
Exclude
button to deactivate these wells for
the current analysis.
Below are definitions of an experimental data set for the different curve fit types:
Linear Standard Curve:
An experimental data set consists of standard curve replicates (i.e., wells with
the well type Known all with the same name and individual concentrations), Blanks and Unknowns (with
either individual names and dilution factors, or replicates with same name and dilution factors).
Linear Response and Dose Response:
These analysis types will allow only one set of experimental curve
replicates to be analyzed at a time. An experimental data set consists of wells with the well type Known (all
with the same name and individual concentrations) and Blanks.
f. Select all of the wells that define an experimental data set, and touch the
Well Grouping
button to
automatically assign replicates based on well type, well name and concentration. After well grouping, all
wells outside of the currently selected experimental data set will be excluded from analysis.
