IGEM:Harvard/2007/Protocols/Magnetic Separation

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  1. Attach the MiniMACS Separation Unit to the MACS MultiStand (up to 4)
  2. Place the MS column in the MiniMACS Separation Unit.
  3. Prepare MS Column b rinsing with buffer: apply 500 ul of buffer on top of the column. Let the buffer run through and discard the effluent.
  4. Apply cell suspension onto the prepared column. Collect unlabeled cells that pass through. Wash column with 3x500 ul of buffer, only adding new buffer once the column reservoir is empty. Collect total effluent. THis is the unlabeled cell fraction.
  5. Remove MS Column from the separator and place it on a suitable collection tube.
  6. Pipette 1 ml of buffer onto the MS Column. Immediately flush out fraction with the magnetically labeled cells by firmly pushing the plunger into the column.

Additional steps:

  1. Apply the positive fraction directly on a new, freshly prepared MS Column.
  2. Wash with 3x500 ul of buffer to remove remaining unlabeled cells.
  3. Remove column from the separator and elute positive fraction in a small buffer volume.
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