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  • Clean Up of Amplified Library (E6285)

    Protocol

    1. Add 90 μl (0.9X 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 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. While the tube is on the magnet, add 200 μl of freshly prepared 80% ethanol.

    6. Incubate at room temperature for 30 seconds and carefully remove and discard the supernatent.

    7. Repeat steps 5-6.

    8. Keeping the tube in the magnetic rack, with the cap open, air dry the beads for 5 minutes.
      Caution: Do not overdry the beads. This may result in lower recovery of DNA target.

    9. Remove the tube from the magnet. Resuspend the beads in 25 μl of 0.1X TE. Mix well on a vortexer or by pipetting up and down, and incubate for 2 minutes at room temperature.

    10. Put the tube on the magnetic rack until the solution is clear. Transfer approximately 20 μl to a clean tube.

    11. Dilute 2-3 μl of the library 1:4 in 0.1X TE. Assess library quality on the Bioanalyzer.