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==Post-Extension SPRI (07/15/2015)== * Vortex AMPure XP beads to resuspend. * Add 101 μl resuspended AMPure XP beads beads to the ligation reaction. * Mix well by pipetting up and down at least 10 times or vortex AMPure XP beads 10 seconds to resuspend. * Incubate for 5 minutes at room temperature. * Quickly spin the tube and place it 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 DNA targets (Caution: do not discard beads). * Add 200 μl of 80% freshly prepared ethanol to the tube while in the magnetic stand. * Incubate at room temperature for 30 seconds, and then carefully remove and discard the supernatant. * Repeat Step 5 once, for a total of two washes. * Air the dry beads for 10 minutes while the tube is on the magnetic stand with the lid open. * Elute the DNA target from the beads by adding 15 μl of 10 mM Tris-HCl, pH 8.0 or 0.1X TE. * Note: Be sure not to transfer any beads. Trace amounts of bead carry over may affect the optimal performance of the polymerase used in the PCR master mix (2X) in the subsequent PCR step. * Mix well by pipetting up and down, or on a vortex. After the solution is clear (about 5 minutes), carefully transfer 15 μl supernatant to a new PCR tube.
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