Bio-Rad BioLogic DuoFlow Pathfinder 80 System DuoFlow Chromatography System Starter Kit - Manual (Page 17)

14
Step Number Start (ml) Step
1. 0.0 Collect fractions of size 2.00 ml during entire run
2. 0.0 Isocratic flow with 100% 25 mM Tris-HCI, pH 8.1,
0% 25 mM Tris-HCI, 0.5 M NaCl, pH 8.1
at 4.00 ml/min for 1.0 ml
3. 1.0 Zero Baseline to set UV baseline to 0.0. Select
either UV detector or QuadTec detector.
4. 1.0 Load inject sample, static loop: Inject 0.5 ml
sample at 4.00 ml/min. You will be injecting the
loop size of 50 µl.
5. 1.5 Isocratic flow with 100% 25 mM Tris-HCI, pH 8.1,
0% 25 mM Tris-HCI, 0.5 M NaCl, pH 8.1
at 4.00 ml/min for 0.8 ml
6. 2.3 Linear gradient with 0% to 50% 25 mM Tris,
0.5 M NaCl, pH 8.1 at 4.00 ml/min for 13.0 ml
7. 15.3 Isocratic flow with 0% 25 mM Tris, pH 8.1,
100% 25 mM Tris, 0.5 M NaCl, pH 8.1
at 4.00 ml/min for 2.8 ml
8. 18.1 Isocratic flow with 100% 25 mM Tris, pH 8.1,
0% 25 mM Tris, 0.5 M NaCl, pH 8.1
at 4.00 ml/min for 8.0 ml
9. 26.1 End of protocol
c. When you have finished programming the method, press the toolbar button
RUN. A dialog box will ask you to name the run. Accept the default Run 1
and click the OK button. You will now see the Run screen (see Figure 5).
The Run Screen
a. The toolbar buttons on the left side of the screen enable you to check that
the screen display ranges for UV (see page 8), QuadTec UV/Vis and
conductivity are correctly set and that the gradient pump pressure limits are
appropriate (700 psi high and 20 psi low limit), for the UNO Q1 column.
b. If you have been equilibrating the column while writing the method, you will
notice that the Status Bar is displaying the flow rate and values for UV,
QuadTec UV/Vis, and conductivity detectors. If necessary, you may wish to
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