GE FineLINE 35 oligo Operating Instructions Manual page 21

Packing procedure
Step
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FineLINE 35 oligo column Operating Instructions 28-9649-57 AA
Action
Calculate the column volume (CV) for the dry Primer Support 200
corresponding to the synthesis scale. Use the values from this table:
Primer Support 200 [g]
5
10
20
Note: The exact ratio is 0.200 g Primer Support 200 per ml column volume.
Calculation to determine the column volume:
1.75 × 1.75 × 3.14 × bed height in cm = CV in ml
Add the required weight of dry Primer Support 200 to a beaker.
Make a slurry by adding 1.2 CV of acetonitrile to the beaker, stir gently and
let stand for 2 to 3 minutes to allow Primer Support to swell properly.
Add the slurry to the column, allow Primer Support to settle for 2 to 3
minutes.
Mark the column glass with a pen where the adapter should be in the
column, based on the calculated CV, or use the premiered lines on the
column.
Wet the adapter with 50 to 95% ethanol.
Connect stop plug to the bottom of the column.
Connect a tube to the adapter and insert the other end securely into a
waste vessel.
Insert the adapter. Adjust the column adapter by hand to the level that
corresponds to the calculated volume from the list below.
Make sure that the adapter is in the correct position.
Lock the adapter in place with the mechanical locking device.
Connect a OD 1/8" tube from valve 6 to the tube from the column outlet to
valve 7.
Note: Keep the OD 1/8" tubing as short as possible.
Start UNICORN method instruction PFlow AB (10 bar flow). Continue for
1.5 to 2.0 CV.
Stop the flow.
Start the run.
CV [ml]
25
50
100
Operation 4
Packing the column 4.1
21