SepFast 6.2 mm ID 0.67 ml column
Packing the column：
• Connect each End Plug to each Extension Plug.
• Insert one of the above assemblies into one end of the Column Body. Push the End Plug until it is fully engaged with the shoulder of the Column Body.
• Screw a 10-32 male/luer female connector to the End Plug. Use a syringe pre-filled with 1 – 2 ml of water to fill the column to a level of 0.2 – 0.3 ml.
• Pipette the resin slurry in and suck the bed down with the syringe. Be careful not to dry the bed. If necessary, pipette in more water or more slurry or pipette out more slurry until the packed volume (under suction) reaches the desired level. Note: the packed volume depends on the type of resin. As a guide, the packed volume should be 0.4 – 0.5 ml for agarose-based spherical particles.
• Pipette water in to fully fill the column. Carefully insert the other assembly (to avoid trapping of air bubble). Push it down slowly until the liquid level reaches the thread. Stop pushing and screw a stop plug to seal the top. Then push the top End Plug down until it is fully engaged in the locking mechanism. Note: don’t remove the syringe in this stage.
• Remove the 10-32 male/luer female connector and the syringe. Screw another Stop Plug into the bottom End Plug.
• Depending on the nature of individual resins, this step may be operated to further settle the particles in the bed. Pre-fill a syringe with 10 ml liquid (ideally the same liquid as the one in the column). Insert it into a 10-32 male/luer female connector and purge out any air in the flow path. Remove the top Stop Plug. Attach the pre-filled syringe to the top End Plug (be sure that no air is trapped in the flow path). Remove the bottom Stop Plug. Push through at least 3 ml of liquid under pressure (e.g as fast as possible) by hand. Seal the bottom with a Stop Plug. Disconnect the syringe and then seal the top. This step can also be done by connecting the column to a chromatography system (such as AKTA) to pump liquid through at high speed (e.g. 3-5 ml/min).