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  • Cleanup of Adaptor Ligated DNA (E6285)


    1. Add 72 µl (1.8 x volume) of AMPure XP Beads to the sample and mix by pipetting up and down.

    2. Incubate for 5 minutes at room temperature.

    3. Pulse-spin the tube and place in a magnetic rack for approximately 2-3 minutes until the beads have collected to the wall of the tube and the solution is clear.

    4. Carefully remove and discard the supernatant without disturbing the beads that contain the DNA target.

    5. Keep the tube on the magnet and add 500 µl freshly prepared 80% ethanol. Incubate to room temp for 30 seconds and carefully remove and discard the supernatant.

    6. Repeat step 5.

    7. Pulse-spin the tube, return to the magnet, and remove any residual ethanol with a pipet.

    8. Keeping the tube in the magnetic rack, with the cap open, air dry the beads for 5 minutes at room temperature.

    9. Resuspend the beads in 25 µl of 0.1X TE (volume may be adjusted for specific size selection protocol).

    10. Pulse spin the tube, and place in the magnetic rack until the beads have collected to the side of the tube and the solution is clear

    11. Transfer approximately 20 µl of the supernatant to a clean tube. Be careful not to transfer any beads
    Alternatively, adaptor ligated DNA can be purified on one column.