Step
Start
Number (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, pH 8.1, 0%
25 mM tris, 0.5 M NaCl, pH 8.1 at 4.00 ml/min for
1.0 ml
3.
1.0
Set UV baseline to 0.0
4.
1.0
Static loop: Inject 0.5 ml sample at 4.00 ml/min for
0.1 min
5.
1.5
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 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
3. When you have finished programming the Protocol Editor, click on the
toolbar button RUN. You will see a dialog box asking you to name the
run. For now, type in Run 1 and click the OK button. You will now see
the Run Control Screen (see Figure 5).
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