ÄKTA start System Cue Card, 29-0240-42 AC
5
Prefill the flow path
Cleaning/prefilling the flow 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 buffer A & B inlets)
Washout fractionation tubing
(clean fractionation
tubing)
Column preparation
(prepare/equilibrate a column)
System cleaning
(clean the flow path using 1 M NaOH
and DM water)
The flow path can also be prefilled 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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•
•
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1.
2.
3.
4.
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.
•
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.
•
•
Switch the
Injection valve to Load
position when
prompted by the message below. Tap
Continue
.
•
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.
1.
2.
via Loop
: A sample loop allows the injection of small
sample volumes (25 μl - 5 ml) onto the column. For
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