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  • Cleanup of Adaptor Ligated DNA (E6270)

    Protocol

    1. Add 180 µl (1.8X 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 side of the tube and the solution is clear.

    4. Carefully remove and discard the supernatant without disturbing the beads.

    5. Keep the tube on the magnet and add 500 µl freshly prepared 80% ethanol. Incubate at room temperature, 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 sterile 0.1X TE (volume may be adjusted for specific size selection protocol.)

    10. Pulse-spin the tube and return to the magnet, 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, being careful not to transfer any beads.
    Alternatively, adaptor ligated DNA can be purified on one column.