Clean Up of Amplified Library- NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent (E6825)
- Add 90 μl (0.9X volume) of AMPure XP Beads 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 2–3 minutes until the beads have collected to the side of the tube and the solution is clear.
- Carefully remove and discard the supernatant without disturbing the beads.
- While the tube is on the magnet, add 200 μl of freshly prepared 80% ethanol.
- Incubate at room temperature for 30 seconds and carefully remove and discard the supernatent.
- Repeat steps 5-6.
- 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.
- 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.
- Put the tube on the magnetic rack until the solution is clear. Transfer approximately 20 μl to a clean tube.
- Dilute 2-3 μl of the library 1:4 in 0.1X TE. Assess library quality on the Bioanalyzer.