Bio-Rad BioLogic Signal Import Module (SIM) - Manual (Page 13)

INTRODUCTION SYSTEM OVERVIEW
1-5
1.5 QUICK START PROCEDURE
The general procedure used to create and run a chromatography experiment on a DuoFlow system is
described below.
1. Install the required devices and instruments on the system (see Figures 3-4 and 5-5 for cable
connections and 4-1 and 4-3 for plumbing connections).
2. Flush all plumbing with DDI H
2
O to ensure that the system is clean and free of air and then prime
the pumps with starting buffer. See Chapter 4 for more detail.
3. Attach a column and set the pressure limits in the Manual screen. The high limit should be less than
or equal to the pressure limit for the column.
4. Equilibrate the column and system with starting buffer. System equilibration is controlled from the
Manual screen (see Section 7.1).
5. Create a new method in the Browser.
a.
Start the Browser using the Browser button on the tool bar and then select or create a user
and project. Refer to Chapter 6 for more information on the Browser screen.
b. Use the New or New/New User option in the Browser tools, on the left side of the screen, to
create a new user and enter a username.
c. Use the New and New Method option in the Browser tools to create and name a new method.
Click OK to proceed to the hardware Setup screen. Alternatively, check the Use Method
Templates box, select a method template and press OK.
6. In the Device Setup screen, select the devices that are connected to the system. Select the
File/Save Setup menu item and save the device setup (check the Default Setup box if this is the
default setup). Refer to Section 7.2 for more information about the Device Setup screen.
7. Start the Protocol Editor using the tool bar Protocol Editor button. Use the protocol screen Add Step
tools to create a new method as illustrated with below for an ion exchange protocol. See Chapter 7
for more information.
a. Add
an Isocratic Flow step to equilibrate the column and enter the required flow rate, step
size and %B. The parameters entered here will automatically appear in the next step, but can
be changed at any time.
b. Add a Zero Baseline step to zero the selected detector prior to sample injection.
c. Add a Load Inject Sample step and then enter the sample inject volume and flow rate. If a
static injection loop is used with an AVR7-3 Sample Inject Valve select static loop. For
information about the other injection options see Chapter 8.
d. Add an Isocratic Flow step to wash the column and enter the required flow rate, step size and
%B.
e. Add a Linear Gradient step to elute the column and enter the required flow rate, step size,
initial %B and final %B.
f. Add an Isocratic Flow step to clean the column and enter the required flow rate step size and
%B (usually 100%). This ensures that the entire sample is removed from the column.
g. Add an Isocratic Flow step to re-equilibrate the column and enter the required flow rate step
size and %B (see step a, above).
h. Add a Fraction Collection step to specify how fractions are to be collected for the experiment.
Enter the collection technique (e.g. collect all, threshold, etc), fraction size and any required
threshold or collection windows parameters. Note that the dialog displays the number of tubes
required for the currently defined protocol.
8. Press the New Run button to create a new run, enter a run name and open the Run screen.
9. Set the appropriate pressure limits for the column being used and then press Start on the system
tool bar. Refer to Chapter 7.4 for more information about the Run screen.