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  • Clean Up of Amplified Library (E6270)

    Protocol

    1. Add 100 µl (1.0X volume) of AMPure XP Reagent 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 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.

    5. Keep the tube on the magnet and add 500 µl freshly prepared 80% ethanol. Incubate 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 35 µl of 0.1 x TE.

    10. Pulse-spin the tube, return to the magnet until the beads have collected to the wall of the tube and solution is clear. 

    11. Transfer approximately 30 µl of supernatant to a fresh tube. Be careful not to transfer any beads.

    12. Assess the library quality on a Bioanalyzer
    Alternatively, adaptor ligated DNA can be purified on one column, elute in 30 µl of 0.1X TE.