Clean Up of Amplified Library - NEBNext Fast DNA Library Prep Set for Ion Torrent (E6270)
- Add 90 µl (0.9X volume) of AMPure XP Reagent to the sample and mix by pipetting up and down.
- Incubate for 5 minutes at room temperature.
- 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.
- Carefully remove and discard the supernatant without disturbing the beads.
- Keep the tube on the magnet and add 200 µl freshly prepared 80% ethanol. Incubate 30 seconds, and carefully remove and discard the supernatant.
- Repeat step 5.
- Keeping the tube in the magnetic rack, with the cap open, air dry the beads for 5 minutes at room temperature.
Caution: Do not overdry the beads. This may result in lower recovery of DNA target.
- Remove the tube from the magnet. Resuspend the beads in 35 μl of 0.1X TE and incubate for 2 minutes at room temperature.
- Pulse-spin the tube, return to the magnet until the beads have collected to the wall of the tube and solution is clear.
- Transfer approximately 30 µl of supernatant to a fresh tube. Be careful not to transfer any beads.
- Dilute 2-3 μl of the library 1:4 in 0.1X TE. Assess the library quality on a Bioanalyzer.