Cleaning A Clogged Column (NOT FPLC Column)


 * 1) Transfer beads to new column.

a.  Check that new column flows well by filling it with diH2O. The column should be a continuous stream when full and be a rapid drip at half full.

b. Transfer beads by resuspending in PBS and transfer PBS/beads slurry to new column. Resuspend/transfer in as little volume at a single time.

c.  Continue to resuspend and transfer small amounts of PBS/bead slurry until all beads are transfered.


 * 1) Wash column with diH2O until residual beads can no longer be seen.  Feel free to cap and shake the column to agitate beads stuck to the walls.
 * 2) Attach the tip of the column to the diH2O spout.  The fit should be snug. Slowly turn on the diH2O tap until a steady stream comes out the top of the column.  Continue flushing in this direction for a minute or so.
 * 3) Attach column to stand with a waste beaker underneath.  Apply 1X column volume of 10% SDS. Fill the column again and cap.  Incubate SDS on column for 5 minutes then let it flow out.
 * 4) Column with water until no more bubbles can be seen.
 * 5) Apply 2 column volumes of .5 M NaOH as in Step 4.  Then wash as in step 5.
 * 6) Test column speed as in Step 1a.  Repeat steps 3-6 as many times as it needed.