Bio-Rad BioLogic Signal Import Module (SIM) - Manual (Page 180)

SAMPLE LOADING ADVANCED SYSTEM APPLICATIONS
8-7
Writing the Protocol
Before running a protocol which involves the use of a DynaLoop, be sure to purge air from the lines, as
discussed in the DynaLoop Instruction Manual, Chapter 2, Installation. Then follow the guidelines which
apply to your application.
The following steps should be programmed within the method protocol to manually load a sample:
1. Ensure that the column is properly equilibrated by using an Isocratic Flow step.
2. From the Manual screen, set the AVR7-3 sample inject valve to its Inject (I) position and allow the
sliding seal to contact the sample end fitting on the DynaLoop using the purge procedure discussed
in the Dynaloop Instruction Manual. Then set the valve to the Load (L) position.
3. Load sample into the DynaLoop by using a syringe. As sample is loaded into the loop, it displaces
the buffer that was used to purge the unit. This design means the DynaLoop cannot be overfilled.
When sufficient volume of sample for a run or series of runs is loaded into the DynaLoop, stop the
filling process. For a series of partial volume injections after one loading sequence, fill the DynaLoop
with about 2 percent extra sample. Leave the syringe in the Inject port to minimize the introduction
of air into the DynaLoop.
The sample is now ready for injection, and the protocol can be programmed.
4. In the Protocol screen, select Load/Inject Sample. In the Load/Inject Sample window, note that the
AVR7-3 valve will automatically move to the “Inject” position at the start of the step and to the “Load”
position at the end of the step.
a. In the Load/Inject Sample window, specify Dynamic Loop.
b. In the “Injection Buffer” area of the window, select the buffer(s) and percent (%) composition to
be used to inject the sample onto the column.
c. In the “Volume (ml)” field, enter the sample volume.
d. In the “Flow (ml/min),” enter the flow rate of the Workstation pump that will be used to inject the
sample onto the column.
5. Continue writing the separation protocol.
The following steps should be programmed within the method protocol to automatically inject a sample using
an auxiliary pump:
1. Connect the Aux pump. The EGP pump connects to the Instrument bus. If you are using the EP-1
Econo pump, consult Section 8.2.
2. In the Setup screen, select Aux Load Pump and AVR7-3 valve in addition to the other device and
instruments you have connected to the system.
3. In the Protocol screen, program an Isocratic Flow step that is long enough for the auxiliary pump to
load the desired quantity of sample into the DynaLoop. For example, if you are loading 25 ml of
sample at an auxiliary pump flow rate of 5 ml/min, you will need at least a 5 minute step prior to the
sample loading step. Otherwise the protocol will fail the automatic validation done prior to the run.
The flow rate of the DuoFlow pump is not critical, so you may set a low flow rate (e.g., 0.1 ml/min) to
minimize buffer waste. It is important that either the time or volume length of this first step is of
sufficient duration to allow the auxiliary pump to fill the DynaLoop.
4. In the Protocol screen, select Load/Inject Sample to program the DuoFlow system to automatically
fill and inject the DynaLoop sample. From the Load/Inject Sample window,
a. Select the Dynamic Loop as the type of loop to be used.