4. Open the switchCONTROL software.
5. Load the biochip, align and prime the system with buffer.
6. Set the temperature of the autosampler tray and chip if
necessary.
7. In the “Taskflow” window, open the folder and add the saved
assay from step 12 (Subheading 3.2). The “Name” and “Project” windows can be filled in order to classify and identify data
files more easily.
8. Start the experiment by clicking on the “Start/stop” button.
3.4 switchSENSE
®
Data Analysis
1. Open the switchANALYSIS software and open a “NEW”
analysis.
2. Load the appropriate dataset of the whole experiment.
3. In this dataset, select the kinetic data.
4. Drag and drop these kinetic data in the right part of the
window.
5. Click on “CREATE NEW ANALYSIS” in the bottom right
part of the window. A new window opens up.
6. Select “Kinetics.” All kinetic data are shown on the left part with all
association and dissociation events being listed as successive blocks
(one block corresponding to one concentration of 80S, Fig. 2).
7. The blank experiment for association and dissociation should
be subtracted by dragging and dropping blank experiments
(0 M concentrations) on the corresponding association or
dissociation blocks for each 80S concentration. This results in
the addition of a new line in the bottom of each block (left part
of the window), and the displayed curve is now corrected from
background.
8. Remove all non-corrected data from the right part of the
window by clicking on the garbage icon on the right.
9. Click on “AUTO-LOAD” in the left bottom part of the window. All background-subtracted data are now displayed on the
main right window (Fig. 3).
10. If necessary, manually select the proper window for data analysis for each 80S concentration in order to remove artifacts
generated by air bubbles following the beginning of each
injection.
11. Click on “FIT ANALYSIS” on the bottom right part of the
window. Results are shown in the main right window (Fig. 4).
12. A publication-quality image can be made by clicking on the
camera icon in the top right part of the window. Results will be
copied in the clipboard as a PNG file. Raw datasets could also
be exported by clicking on the “EXPORT” icon in the bottom
right part of the window.
346
Emma Schenckbecher et al.
5. Load the biochip, align and prime the system with buffer.
6. Set the temperature of the autosampler tray and chip if
necessary.
7. In the “Taskflow” window, open the folder and add the saved
assay from step 12 (Subheading 3.2). The “Name” and “Project” windows can be filled in order to classify and identify data
files more easily.
8. Start the experiment by clicking on the “Start/stop” button.
3.4 switchSENSE
®
Data Analysis
1. Open the switchANALYSIS software and open a “NEW”
analysis.
2. Load the appropriate dataset of the whole experiment.
3. In this dataset, select the kinetic data.
4. Drag and drop these kinetic data in the right part of the
window.
5. Click on “CREATE NEW ANALYSIS” in the bottom right
part of the window. A new window opens up.
6. Select “Kinetics.” All kinetic data are shown on the left part with all
association and dissociation events being listed as successive blocks
(one block corresponding to one concentration of 80S, Fig. 2).
7. The blank experiment for association and dissociation should
be subtracted by dragging and dropping blank experiments
(0 M concentrations) on the corresponding association or
dissociation blocks for each 80S concentration. This results in
the addition of a new line in the bottom of each block (left part
of the window), and the displayed curve is now corrected from
background.
8. Remove all non-corrected data from the right part of the
window by clicking on the garbage icon on the right.
9. Click on “AUTO-LOAD” in the left bottom part of the window. All background-subtracted data are now displayed on the
main right window (Fig. 3).
10. If necessary, manually select the proper window for data analysis for each 80S concentration in order to remove artifacts
generated by air bubbles following the beginning of each
injection.
11. Click on “FIT ANALYSIS” on the bottom right part of the
window. Results are shown in the main right window (Fig. 4).
12. A publication-quality image can be made by clicking on the
camera icon in the top right part of the window. Results will be
copied in the clipboard as a PNG file. Raw datasets could also
be exported by clicking on the “EXPORT” icon in the bottom
right part of the window.
346
Emma Schenckbecher et al.