Bio-Rad BioLogic Signal Import Module (SIM) - Manual (Page 178)

SAMPLE LOADING ADVANCED SYSTEM APPLICATIONS
8-5
Figure 8-4. Sample Loading Through the Workstation Pump
Writing the Gradient Pump Direct Inject Protocol
The protocol Load/Inject step should be programmed as follows:
1. In the protocol Load/Inject step, select Direct Inject as the injection type, select the Pump A (or
Pump B) option, select a sample and set the sample volume and flow rate.
2. Add an isocratic step after the load step to ensure that the entire sample is loaded onto the column.
The inlet tubing and mixer should be rinsed with at least 5 volumes of running buffer to ensure that
the entire sample is loaded onto the column.
3. Add protocol cleaning steps to ensure that all the residual protein has been removed from the tubing
and mixer.
a. Add a Change Valve step to move the AVR7-3 sample inject valve to the Purge position
b. Add an isocratic flow step and run 3-5 volumes of Buffer B through pump B.
c. Add an isocratic flow step and run 3-5 volumes of Buffer A through pump A
d. Add a change Valve step to move the AVR7-3 sample inject valve back to the Load position.
System Sanitization
For sanitization, 1.0 M sodium hydroxide (NaOH) may be used. Be sure to wash with water to remove the
base. Refer to Chapter 11, Maintenance, for information on how to store the DuoFlow system.
A B
RUNNING
BUFFER
SANITIZING SOLUTION
(I.E., 1M SODIUM
HYDROXIDE)
SAMPLE
ELUTION
BUFFER
TO COLUMN
WORKSTATION
STORAGE SOLUTION
(I.E., 20% ETHANOL)
SV5-4