Technical data

Sample Preparation (3 µg DNA Samples) 2
Step 7. Purify the sample using AMPure XP beads
SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing 31
Step 7. Purify the sample using 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 dA-tailed
DNA sample in the PCR plate. Pipette up and down 10 times 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 to 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 15 µL nuclease-free water to each sample well.
13 Seal the wells with strip caps, then mix well on a vortex mixer and
briefly spin the plate 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 13 µL of the cleared supernatant to a fresh PCR plate well. You
can discard the beads at this time.
17 Proceed immediately to the next step, Step 8. Ligate the paired-end
adaptor.