Cleanup of Adaptor Ligated DNA - NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent (E6285)
- Add 72 µl (1.8 x 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 approximately 2-3 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 that contain the DNA target.
- Keep the tube on the magnet and add 200 µl freshly prepared 80% ethanol. Incubate to room temp for 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 25 μl of 0.1X TE (volume may be adjusted for specific size selection protocol). Incubate for 2 minutes at room temperature.
- Pulse spin the tube, and place in the magnetic rack until the beads have collected to the side of the tube and the solution is clear.
- Transfer approximately 20 μl of the supernatant to a clean tube. Be careful not to transfer any beads.
- For E-Gel or Agarose gel size selection, select adaptor ligated DNA in the 190–230 bp range for 100 bp libraries and 290–330 bp range for 200 bp read lengths.
- Proceed to PCR Amplification of Adaptor Ligated DNA.