- For use with AMPure XP beads, allow the beads to warm to room temperature for at least 30 minutes prior to use.
- Vortex beads to resuspend.
- Add 108 μl (1.8X) of resuspended beads to the ligation reaction. Mix thoroughly on a vortex mixer or by pipetting up and down at least 10 times.
- Incubate for 5 minutes at room temperature.
- Put the tube/PCR plate on an appropriate magnetic stand to separate beads from supernatant. After the solution is clear (about 5 minutes), carefully remove and discard the supernatant. Be careful not to disturb the beads that contain the DNA targets.
- Add 200 μl of 80% freshly prepared ethanol to the tube/PCR plate while in the magnetic stand. Incubate at room temperature for 30 seconds, and then carefully remove and discard the supernatant.
- Repeat Step 6 once.
- Air dry beads for 5 minutes while the tube/PCR plate is on the magnetic stand with the lid open.
Caution: Do not overdry the beads. This may result in lower recovery of DNA target. - Remove the tube/plate from the magnet. Elute the DNA target from the beads by adding 47 μl of 10 mM Tris-HCl or 0.1X TE.
- Mix well on a vortex mixer or by pipetting up and down and incubate for 2 minutes at room temperature.
- Put the tube/PCR plate in the magnetic stand until the solution is clear.Without disturbing the bead pellet, carefully transfer 42 μl of the supernatant to a fresh, sterile microfuge tube