8.2 Post Fragmentation, End Repair & A-tailing Double Sided Size Selection – SPRIselect

  1. Vortex to resuspend SPRIselect reagent. Add 30 μl SPRIselect (0.6X) reagent to each sample. Pipette mix 15x (pipette set to 75 μl).

  2. Incubate 5 min at room temperature.

  3. Place on the magnet•High until the solution clears. DO NOT discard supernatant.

  4. Transfer 75 μl supernatant to a new tube strip.

  5. Vortex to resuspend SPRIselect reagent. Add 10 μl SPRIselect reagent (0.8X) to each sample. Pipette mix 15x (pipette set to 80 μl).

  6. Incubate 5 min at room temperature.

  7. Place on the magnet•High until the solution clears.

  8. Remove 80 μl supernatant. DO NOT discard any beads.

  9. Add 125 μl 80% ethanol to the pellet. Wait 30 sec.

  10. Remove the ethanol.

  11. Repeat steps i and j for a total of 2 washes.

  12. Centrifuge briefly. Place on the magnet•Low until the solution clears. Remove remaining ethanol. DO NOT dry.

  13. Remove from the magnet and immediately add 50.5 μl Buffer EB to each sample. Pipette mix 15x.

  14. Incubate 2 min at room temperature.

  15. Place on the magnet•High until the solution clears.

  16. Transfer 50 μl sample to a new tube strip.