ÄKTA start System Cue Card, 29-0240-42 AC 5
Prell the ow path
Cleaning/prelling the ow path including all inlets and
outlets is performed using the system methods listed below.
For instructions on how to prime the sample inlet tubing
and sample loop, refer to ÄKTA start Operating Instructions,
section 5.7 Sample application. For instructions on how to
perform system methods, refer to ÄKTA start Maintenance
Cue Card.
Pump wash A/Pump wash B (prime buer A & B inlets)
Washout fractionation tubing (clean fractionation
tubing)
Column preparation (prepare/equilibrate a column)
System cleaning (clean the ow path using 1 M NaOH
and DM water)
The ow path can also be prelled by using a manual run.
Connect a column
For detailed instructions on how to connect a column, refer
to ÄKTA start Operating Instructions.
Instruction
Depending on column dimension, choose the
appropriate location to place the column. The column
holder rails are indicated in the image below.
Attach an appropriate column holder to the column
holder rail on the instrument.
Mount the column on the union connector if the column
type requires a union.
Fix the column to the column holder, then connect the
tubing to the top and to the bottom of the column.
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Sample application
The sample can be applied to the column in two ways:
via Pump: The sample can be applied directly from a
sample container through the Sample valve as shown
in the image below. It allows application of large volume
of sample (> 5 ml) and facilitates unattended operation.
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larger sample volumes (10 - 150 ml), use a Superloop™.
The loop has to be connected to the Injection valve
(ports 2 and 5) as depicted below.
To apply the sample from a loop:
Fill the loop with sample through port 3 of the Injection
valve (in Load position).
Switch the Injection valve to Inject position when
prompted by the message below. Tap Continue. The
sample is loaded onto the column only when the
Injection valve is in Inject position during the run.
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Switch the Injection valve to Load position when
prompted by the message below. Tap Continue.
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Note: The system is in hold state while injecting the sample
from loop. To ensure that the injection mark coincides with
the injection event, acknowledge the message immediately
after the action is performed.
Note: For AC/IEX methods, when loading the sample
through a loop it is advisable to empty the loop with 3× the
loop volume to achieve good sample recovery.
Cold room operations
Preparation
Place the ÄKTA start instrument in the cold room.
Allow the instrument to stabilize at the temperature of
the cold room.
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via Loop: A sample loop allows the injection of small
sample volumes (25 μl - 5 ml) onto the column. For
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