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  • Agencourt AMPure XP Bead Clean-up and Size Selection of End Repaired DNA (E6260)

    Protocol

    1. Warm the Ampure beads to room temperature and mix thoroughly before use.
    2. Prepare 3.6 mls of 70% ethanol for each sample. The 70% ethanol solution should be prepared fresh.
    3. Add 70 μl of Ampure beads to each sample, mix thoroughly and rotate for 10 minutes at room temperature.
    4. Place samples on a magnetic separator. When the beads have collected to the wall of the tube and the solution is clear, transfer the liquid to a new tube. Be careful not to disturb the beads. The liquid contains the end repaired DNA.
    5. Add 110 μl of Ampure beads to each sample, mix and rotate for 10 minutes at room temperature.
    6. Place the samples on a magnetic separator, when the beads have collected to the wall of the tube and the solution is clear, remove and discard the liquid. The end repaired DNA is now bound to the beads.
    7. Add 300 μl of 70% ethanol. Wash the beads by turning the tube 180° and allowing the beads to re-collect on the side of the tube. Turn the tube 6 times.
    8. Remove and discard the ethanol.
    9. Repeat steps 7 and 8 two more times.
    10. Remove the tubes from the magnetic separator, quick spin the beads, place back on the magnet and remove any remaining liquid. The quick spin will aid in drying the beads.
    11. Keeping the tubes on the magnet and the caps open, dry the beads at room temperature for 20-30 minutes. Cracks will be observed in the bead pellet when drying is complete.
    12. Add 30 μl of water to the dried beads and vortex to mix thoroughly.
    13. Place the samples on a magnetic separator, when the beads have collected to the wall of the tube and the solution is clear, transfer the liquid to a fresh tube. The liquid contains your purified library.