Bio-Rad Media Sampler Pack - Manual (Page 6)

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Section 2. Column Packing
2.1 Recommended Column Packing Buffer System
A solution of 200 mM dibasic sodium phosphate, pH 9–10, is recommended for
packing CFT ceramic fluoroapatite. Historically, solutions up to 400 mM sodium
phosphate have been used, but there is no added benefit from using these higher
concentrations.
2.2 Small Column Packing — Slurry Packing
Slurry packing is preferred for small columns. Use this method for packing 40 µm
CFT ceramic fluoroapatite into 5–50 mm ID columns. For best results, a bed height
of 5–30 cm should be used.
1. Use a column that is 2.5 times longer than the required bed height. Close the
outlet valve.
2. Make a 50% (v/v) slurry of CFT in degassed 200 mM phosphate buffer,
pH 9–10. Suspend the CFT by gently swirling or stirring with a plastic rod. Do
not use a magnetic stirrer.
3. Fill the column to about 10% of its volume with buffer. Ensure that the bottom
bed support is fully saturated and free of bubbles.
4. Pour the slurry into the column and insert the top flow adaptor. Make sure the
tubing is open to allow buffer to flow out through the top adaptor along with
any air as the adaptor is inserted into the column tube.
5. Secure the top adaptor and attach the buffer line from the pump.
6. Open the column outlet and pump buffer through the column at
~400–600 cm/hr for 5–10 min or at the maximum pressure allowed by the
column hardware.
7. Turn off the pump and close the column outlet. Open the tubing of the top flow
adaptor again to allow liquid to exit the column as the flow adaptor is lowered
to rest firmly against the top of the CFT bed.
8. Repeat cycles of flow followed by adjustment of the flow adaptor until the CFT
bed height stops decreasing with flow.
9. Equilibrate.
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