Clean Up Library

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• IPB2 (Illumina Purification Beads 2)
• IDTE Buffer
• Absolute ethanol (EtOH)
• Nuclease-free water (customer provided, sterile)
• 0.2 ml 8-tube strip
1. Prepare the following consumables:
• IPB2—Vortex to resuspend.
• For each reaction, prepare 400 μl fresh 85% EtOH solution from absolute EtOH.
1. If you are resuming after a safe stopping point, briefly centrifuge the sample tube.
2. Add 45 μl nuclease-free water.
3. For 95 μl reaction volumes, add 76 μl IPB2. If needed, adjust the magnetic bead volume for a 0.8X ratio of magnetic beads.
4. Pipette 15 times at the 160 µl stroke to mix.
5. If any magnetic beads remain on the lid or side of the tube, centrifuge briefly.
6. Incubate at room temperature for 5 minutes.
7. Place on the magnetic stand and wait until the liquid is clear (~5 minutes).
8. Without disturbing the beads, remove and discard all supernatant from each tube.
9. Wash beads as follows.
a. Add 200 μl fresh 85% EtOH.
b. Wait 30 seconds.
c. Without disturbing the pellet, remove and discard EtOH.
10. Wash the beads a second time.
11. Without disturbing the beads, use a 20 µl pipette to remove residual EtOH.
12. Air-dry until a slight gloss remains on the bead surface (~2 minutes).

Depending on humidity level, air-drying can take up to 5 minutes.

Overdrying the beads can result in cracks and decrease elution efficiency.

13. Remove from the magnetic stand.
14. Add 21 μl IDTE Buffer.
15. Pipette 10 times at the 20 µl stroke to resuspend.
16. Incubate at room temperature for 5 minutes.
17. Place on the magnetic stand and wait until the liquid is clear (~2 minutes).
18. Transfer 20 μl supernatant to a new PCR tube strip.

SAFE STOPPING POINT

If you are stopping, seal the tubes and store at ‑25°C to ‑15°C for up to 30 days.