Technical data
3 Sample Preparation (200 ng DNA Samples)
Step 10. Purify the amplified library with AMPure XP beads
56 SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing
Step 10. Purify the amplified library with AMPure XP beads
1 Let the AMPure XP beads come to room temperature for at least
30 minutes. Do not freeze the beads at any time.
2 Mix the bead suspension well so that the reagent appears homogeneous
and consistent in color.
3 Add 90 µL of homogeneous AMPure XP beads to each 50-µL amplified
DNA sample in the PCR plate. Pipette up and down to mix.
4 Incubate samples for 5 minutes at room temperature.
5 Put the plate into a magnetic separation device. Wait for the solution to
clear (approximately 3 to 5 minutes).
6 Keep the plate in the magnetic stand. Carefully remove and discard the
cleared solution from each well. Do not touch the beads while removing
the solution.
7 Continue to keep the plate in the magnetic stand while you dispense
200 µL of freshly-prepared 70% ethanol in each sample well.
8 Wait for 1 minute to allow any disturbed beads to settle, then remove
the ethanol.
9 Repeat step 7 and step 8 step once.
10 Seal the wells with strip caps, then briefly spin the plate to collect the
residual ethanol. Return the plate to the magnetic stand for 30 seconds.
Remove the residual ethanol with a P20 pipette.
11 Dry the samples by placing the unsealed plate on the thermal cycler, set
to hold samples at 37°C, for 1 to 2 minutes or until the residual
ethanol completely evaporates.
12 Add 30 µL nuclease-free water to each sample well.
13 Seal the wells, then mix well on a vortex mixer and briefly spin the
plate in a centrifuge or mini-plate spinner to collect the liquid.
14 Incubate for 2 minutes at room temperature.
15 Put the plate in the magnetic stand and leave for 2 to 3 minutes, until
the solution is clear.
16 Remove the cleared supernatant (approximately 30 µL) to a fresh PCR
plate well. You can discard the beads at this time.
Stopping Point If you do not continue to the next step, seal the plate and store at –20°C.