After packing the FPLC column, connect the tubing from the top of the sample loop to loop E on the FPLC system. Attach the tubing from the bottom of the sample loop to waste two on the FPLC system. Then insert one end of the tubing into the column port of the FPLC system and place the other end in the waste bottle.
Insert the tubing from pump A into filtered double distilled water. Next, open the software. Click on the System Control tab.
Double click on the pump settings and select System Pump Inject Loop. Click on the flow rate settings and set the flow rate to 10 milliliters per minute. Enter 0%in the percent B box.
Then start the flow to flush the sample loop with water to push the sliding seal downwards. Once the sliding seal reaches the bottom of the sample loop, stop the flow. Then switch pump A from double distilled water to the starting buffer.
Connect the tubing from the top of the sample loop to waste two and the tubing from the bottom to loop E.Leave the tubing from the column port in the waste and start the flow again at 10 milliliters per minute.
