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  • Agencourt AMPure XP Bead Clean-up (E2612)

    1. Make sure the Agencourt AMPure XP beads are at room temperature and are thoroughly mixed. 

    2. If your enriched sample volume from the previous section is more than 160 μl, split into two tubes.

    3. Add 1.8X volumes of Agencourt AMPure XP Beads to the sample and mix by pipetting up and down.

    4. Incubate for 5 minutes at room temperature.

    5. Pulse-spin the tube and place in a magnetic rack for at least 5 minutes, until the beads have collected to the wall of the tube and the solution is clear. 

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

    7. Keep the tube on the magnet and add 400 μl of freshly prepared 80% ethanol.

    8. While keeping the tube in the magnetic rack, rotate the tube 90 degrees. When the beads have collected to the opposite side of the tube (near the magnet), turn the tube another 90 degrees. Rotate the tube two more times, each time waiting until the beads collect to the opposite side of the tube.

    9. Allow the solution to become clear, and carefully remove and discard the supernatant. 

    10. Repeat steps 7–9. 

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

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

    13. Resuspend the beads in 50 μl of 0.1X TE Buffer (or use 25 μl for each tube if you split the sample into two tubes in Step 2). 

    14. Pulse-spin the tube, return to the magnet and collect the supernatant (combine the supernatant if you split the sample into two tubes in Step 2).

    15. The sample can now be used for NGS library construction or other downstream analysis.
    Alternatively, enriched samples can be purified by ethanol precipitation.