NEBExpress® Ni Spin Columns Quick Start Protocol (NEB #S1427)

  1. Remove the bottom tab of the column and place the column in a collection tube, loosen the cap.

  2. Centrifuge column at 800 x g for 1 minute, discard the buffer.

  3. Add 250 μl of Lysis/Binding buffer to the column.

  4. Centrifuge column at 800 x g for 1 minute, discard the Lysis/Binding buffer.

  5. Add up to 500 μl of sample lysate to the column, tap to mix and allow binding for 2 minutes.

  6. Centrifuge column at 800 x g for 1 minute, retain flow through.

  7. Place the column in a new 2 ml microcentrifuge tube.

  8. Add 250 μl of Wash buffer to the column and centrifuge at 800 x g for 1 minute, repeat twice. Retain the washes.

  9. Add 200 μl of Elution buffer to the column. Tap the column repeatedly to mix.

  10. Centrifuge at 800 x g for 1 minute, retain the eluate.

  11. Repeat elution step once, collecting fraction in a new microcentrifuge tube.